KR20170003923A - Polysubstituted pyridine compound, preparation method, use and pharmaceutical composition - Google Patents

Abstract

The present invention provides a multisubstituted pyridine compound of general formula I, a preparation method thereof, a use thereof, and a pharmaceutical composition. The multisubstituted pyridine compounds of the general formula I according to the present invention have excellent antitumor effects, while at the same time inhibiting various cell kinases, have excellent pharmacokinetic properties and are well suited for oral and intravenous administration. The pharmaceutical composition according to the present invention may be useful for treating tumors and cancers.

Description

(Polysubstituted pyridine compound, preparation method, use and pharmaceutical composition)

Field of the Invention The present invention relates to the field of pharmacological chemistry, and more particularly to multisubstituted pyridine compounds, a method for producing the same, a use thereof, and a pharmaceutical composition.

Research and development of antitumor drugs in the field of life sciences is the most challenging and important. Recently, with the rapid development of molecular biology and a deeper understanding of the emergence, progression, and mechanism of cancer, the signaling of malignant tumor cells, regulation of the cell cycle, induction of apoptosis, And the interactions between extracellular matrix are progressively elucidated. Accordingly, finding novel anticancer agents with high efficiency, low toxicity and strong specificity that selectively affect a specific target is one of the important fields of pharmaceutical research and development. Thus, it leads to new anticancer drugs - molecular targeted drugs.

Molecular targeting therapeutics refers to a class of drugs that are involved in receptor or key enzymes in transduction that accompany cell cancerization and that inhibit tumor growth at the molecular level. This molecular targeted drug targets the characteristic molecules of tumor cells and acts as an anti-cancer agent by reducing toxicity and side effects to normal cells.

The balance between positive regulators and negative regulators controls tumor angiogenesis, which promotes tumor growth and metastasis, so that the development of angiogenesis inhibitors is a hotspot in tumor research (hotspots). VEGFR is a class of important tyrosine kinases. Many studies have shown that the dysfunction of the signaling pathway of VEGFR plays an important role in tumor development, growth and metastasis. VEGFR mainly comprises VEGFR21 (Flt21), VEGFR22 (KDR / Flt21) and VEGFR23 (Flt24) belonging to the tyrosine kinase receptor. VEGF exerts its biological function by binding to two trans-membrane receptors of endothelial cells.

Signal transduction factors in cell differentiation include many protein kinase families. During cellular signal transduction, protein tyrosine kinase promotes the transfer of phosphate groups from ATP to the tyrosine residues of many key proteins for phosphorylating activation of the by-pass, leading to cell growth, It is very important because it can affect proliferation and differentiation. In many tumor cells, tyrosine kinase activity is abnormally elevated. Over 50% of oncogenes and their products have protein tyrosine kinase activity and abnormal expression can lead to the appearance of tumors. In addition, abnormal expression of the enzyme may be associated with tumor metastasis, tumor angiogenesis, and tumor resistance to chemotherapy. Studies of selective protein kinases that block or alter abnormal signal transduction are thought to be a promising pathway for drug development. Currently, some protein kinase inhibitors and small-molecule therapeutic agents have been found against different ATP-binding sites of protein kinases and have entered clinical studies, such as tyrosine kinase inhibitors.

Sorafenib (trade name: Nexavar), developed by Bayer Pharmaceuticals, was approved by the US Food and Drug Administration (FDA) in December 2005 as the first drug for the treatment of advanced renal carcinoma Is a multi-targeted drug and is the first multi-target drug approved for targeted treatment in clinics around the world. Chinese patent application document CN1341098A discloses the chemical structure of sorapanib, the chemical structure of which is as follows:

Prior to the present invention, another Chinese patent application and patent application document CN102532113A (Application No. 201110435847.9) filed by the present inventor discloses a compound of the following general formula.

Existing anticancer agents still do not meet the therapeutic requirements for the oncological diseases in humans and other mammals and the therapeutic effects of commercially available anticancer agents in clinic treatment still do not reach the desired level, so that more effective anticancer agents are still desired have.

In order to solve the above problems, the present invention provides a multisubstituted pyridine compound, a preparation method thereof, a use thereof, and a pharmaceutical composition.

Especially,

In a first aspect, the present invention provides a multisubstituted pyridine compound of formula I, or a hydrate, solvate or pharmaceutically acceptable salt thereof:

Wherein X < ₁ > is selected from substituted or unsubstituted 5 membered heteroaromatic rings of formula a;

R ₄ , R ₅ and R ₆ are each independently selected from the group consisting of a carbon atom, a nitrogen atom, an oxygen atom or a sulfur atom, and R ₈ , R ₉ and R ₁₀ are each independently selected from the group consisting of hydrogen, halogen, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxyl;

X ₂ is selected from the group consisting of F and H;

X ₃ is halogen, -CN, C ₁ -C ₄ alkyl, halogenated C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxyl, halogenated C ₁ -C ₄ An alkoxyl, and -NR ₁₁ R _12, wherein R ₁₁ And R ₁₂ are each independently selected from the group consisting of hydrogen and C ₁ -C ₄ alkyl.

Preferably, R ₄ , R ₅ and R ₆ are each independently selected from the group consisting of carbon atoms and nitrogen atoms.

Preferably, R ₄ , R ₅ and R ₆ are not simultaneously carbon atoms.

Preferably, R ₄ , R ₅ and R ₆ are not simultaneously nitrogen atoms.

Preferably, R ₈ , R ₉ and R ₁₀ are each independently selected from the group consisting of hydrogen and methyl.

이다.Preferably, X < _{1 >}

to be.

Preferably, X ₃ is _{F, Cl, Br, -CF 3} , -CN, C 1 -C 2 is selected from alkyl, C ₁ -C ₂ alkoxyl, and the group consisting of -NR ₁₁ R _12, wherein R ₁₁ And R ₁₂ are each independently selected from the group consisting of hydrogen and C ₁ -C ₂ alkyl.

Preferably, X < ₃ > is selected from the group consisting of F, Cl and -CN.

Preferably, the multisubstituted pyridine compounds of formula I are selected from the group consisting of:

,

,

,

,

,

,

,

,

,

,

,

,

, 및

, And

More preferably, the multisubstituted pyridine compounds of formula I are selected from the group consisting of the following compounds:

, 및

, And

Preferably, the pharmaceutically acceptable salts of the multisubstituted pyridine compounds of formula I include, but are not limited to, hydrochlorides, hydrobromides, sulfates, phosphates, methanesulfonates, trifluoromethanesulfonates, benzenesulfonates, p- Salts of organic acids such as hydrochloric acid, hydrobromic acid, nitric acid, phosphoric acid, nitric acid, nitric acid, phosphoric acid, nitric acid, Salicylic acid salts, phenylacetic acid salts and mandelic acid salts.

The present invention also relates to a process for the preparation of said multisubstituted pyridine compounds according to the invention, which process comprises the steps of:

1) reacting a compound of formula B and a compound of formula C in the presence of potassium tert-butoxide as a base to give a compound of formula D, as shown by the following scheme:

,

,

Wherein R ₁₃ is F, Cl, Br or I.

2) The compound of formula D and the compound of formula E are reacted in the presence of tetrakis (triphenylphosphine) palladium or bis (triphenylphosphine) palladium (II) dichloride as catalyst, Lt; / RTI > to obtain a compound of general formula F:

; 및

; And

3) reacting a compound of formula F with a compound of formula G to obtain a multisubstituted pyridine compound of formula I, as shown by the following scheme:

Preferably, the compound of formula B is prepared by the following method:

Halogenating the compound of formula A gives the compound of formula B:

Wherein R ₁₃ is F, Cl, Br or I.

Preferably, in the case where X ₃ is NH _2, the manufacturing method including the following steps:

1) reacting a compound of formula H and a compound of formula C as a catalyst in the presence of potassium tert-butoxide to give a compound of formula W, as shown in the following scheme:

;

;

Wherein R < ₁₃ > is F, Cl, Br or I;

2) The compound of formula W and the compound of formula E are reacted in the presence of tetrakis (triphenylphosphine) palladium or bis (triphenylphosphine) palladium (II) dichloride as catalyst, To obtain a compound of the general formula J:

;

;

3) hydrogenating the compound of formula J in the presence of palladium-carbon as catalyst to give the compound of formula K:

; 및

; And

4) reacting a compound of general formula K and a compound of general formula G to obtain a compound of general formula L, as shown by the following scheme:

Here, in the present invention, LDA represents lithium diisopropylamide; THF represents tetrahydrofuran; -78 deg represents -78 deg. C; DMSO represents dimethylsulfoxide; rt represents room temperature; DCM represents dichloromethane; conc. represents "enriched "; TEA represents triethylamine.

In addition, the present invention relates to a method of inhibiting VEGFR-2 (vascular endothelial growth factor receptor-2), VEGFR-3 (vascular endothelial growth factor receptor-3), CRAF (human C-RAF native carcinoma / threonine protein kinase BRAF, V600E BRAF, KIT, and / or FLT-3 (FMS type tyrosine kinase 3), BRAF (human serine / threonine protein kinase), PDGFR- Or a hydrate, solvate or pharmaceutically acceptable salt thereof according to the present invention in the manufacture of a medicament for the treatment and / or prophylaxis of diseases associated with kinase.

In an embodiment of the present invention, the diseases associated with VEGFR-2, VEGFR-3, CRAF, PDGFR-beta, BRAF, V600E BRAF, KIT and / or FLT-3 kinase include tumors or cancers.

Preferably, the tumor or cancer is selected from the group consisting of malignant melanoma, liver cancer, renal carcinoma, acute leukemia, chronic leukemia, non-small cell lung cancer, small cell lung cancer, prostatic cancer, thyroid cancer, skin cancer, colon cancer, rectal cancer, pancreatic cancer, ovarian cancer, Breast cancer, mammary cancer, myelodysplastic syndromes, esophageal cancer, or mesothelioma.

The present invention also provides a method for the treatment and / or prophylaxis of diseases associated with VEGFR-2, VEGFR-3, CRAF, PDGFR-beta, BRAF, V600E BRAF, KIT and / or FLT- There is provided a method comprising administering to a subject a therapeutically or prophylactically effective amount of said multisubstituted pyridine compound, or a hydrate, solvate or pharmaceutically acceptable salt thereof, according to the invention.

In an embodiment of the present invention, the diseases associated with VEGFR-2, VEGFR-3, CRAF, PDGFR-beta, BRAF, V600E BRAF, KIT and / or FLT-3 kinase include tumors or cancers.

Preferably, the tumor or cancer is selected from the group consisting of malignant melanoma, liver cancer, kidney cancer, acute leukemia, chronic leukemia, non-small cell lung cancer, prostate cancer, thyroid cancer, skin cancer, colon cancer, pancreatic cancer, ovarian cancer, breast cancer, Syndrome, esophageal cancer, or mesothelioma.

The present invention also provides a method for the treatment and / or prophylaxis of diseases associated with VEGFR-2, VEGFR-3, CRAF, PDGFR-β, BRAF, V600E BRAF, KIT and / Solvates, solvates and pharmaceutically acceptable salts thereof.

In an embodiment of the present invention, the disease associated with VEGFR-2, VEGFR-3, CRAF, PDGFR-beta, BRAF, V600E BRAF, KIT and / or FLT-3 kinase is a tumor or cancer.

Preferably, the tumor or cancer is selected from the group consisting of malignant melanoma, liver cancer, kidney cancer, acute leukemia, chronic leukemia, non-small cell lung cancer, prostate cancer, thyroid cancer, skin cancer, colon cancer, pancreatic cancer, ovarian cancer, breast cancer, Syndrome, esophageal cancer, or mesothelioma.

The present invention also relates to a method for inhibiting the activity of VEGFR-2, VEGFR-3, CRAF, PDGFR-β, BRAF, V600E BRAF, KIT and / or FLT-3 kinase in a cell, A pharmaceutically acceptable salt thereof, or a hydrate, solvate and a pharmaceutically acceptable salt thereof, to said cell.

Preferably, the method is carried out in vitro .

Preferably, the method is carried out in vivo.

Preferably, the cell is a cell line, a cell derived from a subject such as a tumor cell or a cancer cell.

Preferably, the tumor or cancer is selected from the group consisting of malignant melanoma, liver cancer, kidney cancer, acute leukemia, chronic leukemia, non-small cell lung cancer, prostate cancer, thyroid cancer, skin cancer, colon cancer, pancreatic cancer, ovarian cancer, breast cancer, Syndrome, esophageal cancer, and mesothelioma.

The present invention also relates to the use of the above-mentioned polysubstituted pyridine compound or its hydrate, solvate or pharmaceutically acceptable salt thereof according to the present invention in the manufacture of a medicament, wherein said medicament is VEGFR-2, VEGFR-3, CRAF, PDGFR-beta, BRAF, V600E BRAF, KIT and / or FLT-3 kinase.

Preferably, the medicament is used in an in vitro method.

Preferably, the agent is used in an in vivo method.

Preferably, the cell is a cell line, or a cell derived from a subject such as a tumor cell or a cancer cell.

Preferably, the tumor or cancer is selected from the group consisting of malignant melanoma, liver cancer, kidney cancer, acute leukemia, chronic leukemia, non-small cell lung cancer, prostate cancer, thyroid cancer, skin cancer, colon cancer, pancreatic cancer, ovarian cancer, breast cancer, Syndrome, esophageal cancer, and mesothelioma.

The present invention also provides a method for inhibiting the activity of VEGFR-2, VEGFR-3, CRAF, PDGFR-beta, BRAF, V600E BRAF, KIT and / or FLT- Substituted pyridine compounds, or hydrates, solvates, and pharmaceutically acceptable salts thereof.

Preferably, it is used in an ex vivo method.

Preferably, it is used in an in vivo method.

Preferably, the cell is a cell line, or a cell derived from an object such as a tumor cell or a cancer cell.

Preferably, the tumor or cancer is selected from the group consisting of malignant melanoma, liver cancer, kidney cancer, acute leukemia, chronic leukemia, non-small cell lung cancer, prostate cancer, thyroid cancer, skin cancer, colon cancer, pancreatic cancer, ovarian cancer, breast cancer, Syndrome, esophageal cancer, and mesothelioma.

The present invention also provides a kit for inhibiting the activity of VEGFR-2, VEGFR-3, CRAF, PDGFR-β, BRAF, V600E BRAF, KIT and / or FLT- Or a hydrate thereof, a solvate or a pharmaceutically acceptable salt thereof, and, optionally, instructions.

In an embodiment of the present invention, the diseases associated with VEGFR-2, VEGFR-3, CRAF, PDGFR-beta, BRAF, V600E BRAF, KIT and / or FLT-3 kinase include tumors or cancers.

Preferably, the tumor or cancer is selected from the group consisting of malignant melanoma, liver cancer, kidney cancer, acute leukemia, chronic leukemia, non-small cell lung cancer, prostate cancer, thyroid cancer, skin cancer, colon cancer, pancreatic cancer, ovarian cancer, breast cancer, Syndrome, esophageal cancer, or mesothelioma.

The present invention also relates to pharmaceutical compositions comprising said polysubstituted pyridine compounds according to the invention, or a hydrate thereof, a solvate thereof and a pharmaceutically acceptable adjuvant (such as a carrier or an excipient) .

Preferably, the pharmaceutical composition is an injection, an oral formulation, a cutaneous permeable agent, or a suppository.

Preferably, the pharmaceutical composition is used for the treatment and / or prevention of diseases associated with VEGFR-2, VEGFR-3, CRAF, PDGFR-beta, BRAF, V600E BRAF, KIT and / or FLT-3 kinase.

In the present invention, C ₁ -C ₄ alkyl is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-butyl.

In the present invention, the C ₁ -C ₄ alkoxyl represents C ₁ -C ₄ alkyl-O-, wherein C ₁ -C ₄ alkyl has the same meaning as defined above.

In the present invention, the halogen is selected from F, Cl, Br, and I.

In the present invention, the C ₁ -C ₂ alkyl represents methyl or ethyl.

In the present invention, the C ₁ -C ₂ alkoxyl represents methoxyl or ethoxyl.

If the name of the compound used in the present specification does not match the formula, the formula may be applied first, or it may be determined by a person skilled in the art according to the practical conditions in combination with common knowledge.

Some of the compounds according to the present invention may be crystallized or recrystallized with water or various organic solvents, in which case various solvates may be formed. The present invention includes stoichiometric solvates containing hydrates and also includes compounds containing variable amounts of water formed when prepared by sublimation under low pressure.

According to the present invention, since the compound of the general formula I is used for pharmaceutical purposes, it is preferred that the compound be in pure form, for example at least 60% purity, most preferably at least 75% % Purity, most preferably at least 98% purity ("%" represents weight%). Impure compounds may be used for the production of the purer forms used in pharmaceutical compositions. The impure product contains at least 1%, more suitably 5%, more preferably 10%, of the compound of general formula I or a pharmaceutically acceptable derivative thereof.

The present invention also relates to a pharmaceutical composition comprising at least one compound of formula I and at least one pharmaceutically acceptable carrier or excipient. The compounds of general formula I or their pharmaceutically acceptable salts may be used alone or in combination with pharmaceutically acceptable carriers or excipients in the form of pharmaceutical compositions. When the compound is used in the form of a pharmaceutical composition, it may be formulated as an aqueous solution or suspension of the compound of formula I, or a pharmaceutically acceptable salt or solvate thereof, and an effective amount of one or more pharmaceutically acceptable carriers or excipients, It is common to prepare an administration form or a dosage form. The process includes mixing, granulating, compressing or dissolving the ingredients by suitable means.

The pharmaceutical composition according to the present invention may be administered by any of the following means: oral administration, spray inhalation, rectal administration, intranasal administration, Vaginal administration, topical administration, intravenous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, Parenteral administration such as intracranial injection or infusion or administration by an explant reservoir such as intravenous injection, intramuscular injection, intrasternal injection, intracranial injection or injection, Administration, intramuscular injection, intraperitoneal administration or intravenous administration are preferred.

Pharmaceutically acceptable carriers included in the pharmaceutical composition of the present invention include serum proteins such as ion exchangers, aluminum oxide, aluminum stearate, lecithin, human serum proteins and the like; A salt or electrolyte of water, a protamine sulfate, etc., a disodium hydrogen phosphate, a potassium hydrogen phosphate, a sodium chloride, a zinc salt, a colloidal silica, a magnesium triacetate, a calcium phosphate, But are not limited to, polyvinylpyrrolidone, cellulose-based materials, polyethylene glycol, carboxymethylcellulose sodium, polyacrylic acid esters, beewax, lanocerin, and the like. For pharmaceutical compositions, the carrier is present in an amount of from 1% to 98% by weight, generally about 80% by weight. For convenience of use, local anesthetics, preservatives, buffering agents and the like may be dissolved directly in the carrier.

Oral formulations such as tablets and capsules for oral use may contain excipients such as syrup, gum arabic, sorbitol, tragacanth or polyvinylpyrrolidone; Fillers such as lactose, sucrose, corn starch, calcium phosphate, sorbitol or aminoacetic acid; Lubricants such as magnesium stearate, talc, polyethylene glycol or silica; Disintegrating agents such as potato starch; Or an acceptable lubrication promoting agent such as sodium lauryl sulfate.

The pharmaceutical compositions of the present invention are in oral liquid form and may be prepared in the form of a suspension, solution, emulsion, syrup or elixir of water and oil, or supplemented with water or other suitable vehicle before use Or as a dry product. Liquid formulations include conventional additives such as suspending agents such as sorbitol, methylcellulose, glucose syrup, gel, hydroxyethylcellulose, carboxymethylcellulose, aluminum stearate gel or hydrogenated edible fats; Emulsifiers such as lecithin, sorbitan monooleate or gum arabic; Or a fat such as almond oil or glycerol, a non-aqueous carrier such as ethylene glycol or ethanol (which may include edible oil); Methyl parahydroxybenzoate, or preservatives such as propyl, sorbic acid and the like.

If necessary, a perfume or a coloring agent may be added. Suppositories may contain conventional suppository bases such as cocoa butter or other glycerides. For parenteral administration, a liquid dosage form will generally consist of a compound and at least one sterile or sterile carrier. The carrier is optimally water. Depending on the selected carrier and the concentration of the drug, the compound may be dissolved in a carrier or prepared as a suspension. In preparing an injection solution, the compound is first dissolved in water, filtered and sterilized and then packaged in a seal bottle or ampoule. When topically administered to the skin, the compound according to the invention may be formulated in a suitable form of ointment, lotion, or cream, wherein the active ingredient is suspended or dissolved in one or more carriers. Carriers for ointment preparation include, but are not limited to, mineral oil, liquid paraffin, albolene, propylene glycol, polyoxyethylene, polyoxypropylene, emulsifying wax and water . The carriers for lotions and creams include, but are not limited to, mineral oil, sorbitan monostearate, Tween 60, hexadecyl ester wax, hexadecane aromatic alcohol, 2-octyldodecanol, benzyl alcohol and water. According to the route of administration, the composition may contain an active ingredient in an amount of 0.1% by weight or more suitably 10-60% by weight. However, when the composition is in unit dosage form, each unit preferably contains 50 to 500 mg of active ingredient. Depending on the route and frequency of administration, the therapeutic dose suitable for an adult is, for example, 100-3000 mg per day, for example 1500 mg per day.

It will be appreciated that the optimal administration dose and interval of the compound of formula I depends on the severity of the disease or disorder, the nature of the compound, the mode of administration, the route of administration and the site of administration, as well as the conditions of the particular mammalian subject being treated shall. The optimal dose may be determined by the physician.

The multisubstituted pyridine compounds according to the present invention have the following advantages and positive effects over the prior art:

 The present invention provides a new class of multisubstituted pyridine compounds for the first time. Compared to the existing compounds (Sorafenib, i.e. the compounds disclosed in CN1341098A or CN102532113A), the multisubstituted pyridine compounds of the general formula I according to the invention have a better antitumor effect, (VEGFR-2), Vascular Endothelial Growth Factor Receptor-3 (VEGFR-3), CRAF, platelet derived growth factor receptor-β (PDGFR-β), BRAF, V600E BRAF, KIT and FLT- 3 < / RTI > kinase, can inhibit various kinases in the cell and on the cell surface simultaneously. In particular, preferred compounds according to the present invention have dual anti-tumor effects. On the other hand, these compounds can block angiogenesis of the tumor by inhibiting VEGFR and PDGFR, thereby inhibiting the growth of tumor cells; On the other hand, these compounds inhibit the growth of tumors by inhibiting the RAF / MEK / ERK signaling pathway, resulting in more effective antitumor effects.

In addition, the compounds according to the invention are notably superior to sorapenib, which is a commercially available drug for in vivo data on blood concentrations and the like, with remarkably excellent anti-tumor effects, as well as excellent pharmacokinetic characteristics , And are well suited for oral and intravenous administration.

In order to obtain a more effective antitumor drug, the present inventors conducted a number of screening tests. For example, we scrambled at positions of substituents X3, X4 and X5 of the pyridine ring of formula II by pharmacodynamic experiments.

The present inventors have surprisingly found by pharmacodynamic experiments that when all of X ₄ and X ₅ in the pyridine ring are hydrogen and X ₃ is substituted with any substituent due to the electron cloud effect and the change in configuration of the compound molecule, The interaction between the atom and the pharmacophoric group 2- (1-methyl-4-pyrazolyl) or 2- (methylcarbamoyl) is improved and the binding strength between the compound molecule and the receptor becomes higher . Further, the present inventors have surprisingly found that when X ₃ is hydrogen in the pyridine ring, the position of X ₃ is a site where the compound can be easily metabolized; The efficacy of the compounds according to the invention by virtue of their ability to block the site of the substituent when X ₃ of the general formula I is substituted by some substituent, thereby improving the metabolic stability of the compound in vivo and ensuring a high blood concentration of the compound To a greater extent. Based on these findings, the present inventors have reached a technical solution of the present invention. The compounds according to the present invention have considerably superior antitumor effects and are significantly better than the compounds disclosed in Chinese patent application CN1341098, CN201110435847.9, as well as sorapenib, a commercially available drug.

The present inventors have also found that the compound according to the present invention had a better therapeutic effect when X ₃ was an electron withdrawing group. Preferred electron withdrawing groups are F, Cl or cyano.

Significant differences between the technical solution of the present invention and the technical solution of the patent application CN201110435847.9 as filed earlier by the present inventors are that the compound according to the invention is present at the 3 position of the pyridine ring The compound disclosed in CN201110435847.9 is substituted in X3, X4 and X5 of the general formula II at the 3, 5 and 6 positions (i.e., the instant specification) of the pyridine ring, It is.

In addition, the commercially available anticancer drug sorapenib is substituted at the 3, 5, and 6 positions of the pyridine ring (i.e., X3, X4, and X5 of Formula II according to the present invention) Are structurally different.

We have found, by pharmacodynamic comparative experiments, that the compounds according to the invention are not substituted at the 3, 5 and 6 positions of the pyrimidine ring (in the specification according to the invention X 3, X 4 and X 5) , CN1341098, CN201110435847.9. ≪ / RTI > In addition, the compound according to the present invention is significantly superior to sorapenib, which is a commercially available anticancer agent, as compared with sorapenib, which is a commercially available anticancer agent, that the compound according to the present invention is a more effective anticancer compound capable of inhibiting a large number of kinases .

The compounds according to the invention have remarkably superior antitumor effects as well as considerably superior pharmacokinetic properties and are clearly superior to sorapenib, a commercially available drug for in vivo data on blood concentrations and the like, for oral and intravenous administration It is very suitable.

Figure 1 shows the half maximal inhibitory concentration of the free base of Example 1, Example 2, Comparative Example 3, Comparative Example 4, and Comparative Example 5 for kinase VEGFR2. will be.
The correspondence between the compounds shown in the figures and the examples is as follows:

The present invention is further described with reference to the following embodiments and drawings. Based on the basic idea of the present invention, those skilled in the art can make various changes or improvements. These changes or improvements are included in the scope of the present invention unless they depart from the basic idea of the present invention.

In the following examples, all reagents, except as otherwise specified, are commercially available from, for example, J & K SCIENTIFIC Co. Ltd., Commercially available from Alfa Aesar (tianjin) Chemical Co. Ltd., or Beijing Ouhe Technology Co., Ltd.

In the following examples, the formula for the yield is:

Yield = product weight x molar mass of raw material / (weight of raw material x molar mass of product)

Example 1

FD-2013015:

3- (4- (3-fluoro-2- (l-methyl-4-pyrazolyl) -pyridin-4-yl-oxy) Phenyl) urea

Manufacturing method:

Step 1: Synthesis of 2-chloro-3-fluoro-4-chloropyridine

(2.4 M hexane solution, 13.13 mL, 31.5 mmol) was added to a solution of diisopropylamine (3.18 g, 31.5 mmol) in anhydrous tetrahydrofuran (30 mL) at -30 캜 under an atmosphere of nitrogen gas. mmol) < / RTI > to give a reaction mixture. The reaction mixture was stirred at -30 占 폚 for 30 minutes, then cooled to -78 占 폚. A solution of 2-chloro-3-fluoropyridine (3.95 g, 30 mmol) in anhydrous tetrahydrofuran (20 mL) was added dropwise and the reaction mixture was then stirred at -78 째 C for 60 minutes. A hexachloroethane solution (7.10 g, 30 mmol) in anhydrous tetrahydrofuran (50 mL) was added dropwise, and then the reaction mixture was stirred at -78 째 C for 60 minutes. The reaction mixture was quenched with a saturated ammonium chloride solution (50 mL), diluted with water (50 mL) and extracted with ethyl acetate (100 mL x 3). Organic phases were combined. The combined organic phases were washed with saline solution (100 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (petroleum ether: ethyl acetate = 100: 1) to give the product (3.60 g, yield: 72%) as a yellow solid.

_{^{1 H NMR (300MHz, CDCl 3}} ): 8.14 (d, J = 5.1Hz, 1H), 7.34 (t, J = 5.1Hz, 1H)

MS (ESI +): m / z 166.2 [M + H] < ^{+ &}

Step 2: Synthesis of 2-chloro-3-fluoro-4- (4-aminophenoxy)

4-Aminophenol (24.8 g, 227 mmol) in anhydrous dimethyl sulfoxide (210 mL) was bubbled with nitrogen gas for 10 min followed by the addition of potassium tert-butoxide (26.80 g, 238.8 mmol) . The reaction mixture was stirred at room temperature for 30 minutes, then 2-chloro-3-fluoro-4-chloropyridine (37.68 g, 227 mmol) was added. The reaction mixture was stirred at room temperature for 5 hours, then diluted with water (100 mL) and extracted with ethyl acetate (500 mL x 3). Combined organic phases. The combined organic phases were washed with brine (500 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 5: 1, v / v) to give the product (15.0 g, yield: 28%) as a pale yellow solid.

^{1 H NMR (300MHz, DMSO-} d 6): δ5.22 (br s, 2H), 6.62 (d, J = 9.0Hz, 2H), 6.75 (t, J = 5.7Hz, 1H), 6.92 (d, J = 9.0 Hz, 2H), 8.05 (d, J = 5.7 Hz, 1H)

MS (ESI +): m / z 239.1 [M + H] < ^{+ &}

Step 3: Synthesis of 4- (3-fluoro-2- (1-methyl-4-pyrazolyl) -pyridin-

To a solution of 2-chloro-3-fluoro-4- (4-aminophenoxy) pyridine (7.2 g, 30.2 mmol), 1-methylpyrazol-4-yl-boronic acid pinacol in tetrahydrofuran (THF, A mixture of ester (6.3 g, 30.2 mmol), potassium carbonate (12.5 g, 90.6 mmol) and tetrakis (triphenylphosphine) palladium (1.74 g, 1.5 mmol) and water (30 mL) , Followed by stirring at 85 캜 for 24 hours under an atmosphere of argon gas to obtain a reaction mixture. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (100 mL x 3). Combined organic phases. The combined organic phases were washed with brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 1: 2, v / v) to give the product (5.4 g, yield: 60%) as a pale yellow solid.

^{1 H NMR (300MHz, DMSO-} d 6): δ3.93 (s, 3H), 5.18 (br s, 2H), 6.54 (t, J = 5.7Hz, 1H), 6.63 (d, J = 8.7Hz, J = 5.7 Hz, 2H), 6.91 (d, J = 8.7 Hz, 2H), 7.98 (d, J = 0.6 Hz, 1H)

MS (ESI +): m / z 285.1 [M + H] < ^{+ &}

Step 4: l- (4-Chloro-3- (trifluoromethyl) phenyl) -3- (4- (3-fluoro- -Yl-oxy) phenyl) urea: < EMI ID =

To a solution of 4- (3-fluoro-2- (l-methyl-pyrazol-4-yl) -pyridin-4-yl-oxy) aniline (1.71 g, 6.0 mmol) -3-trifluoromethylphenyl isocyanate (1.6 g, 7.2 mmol) was stirred at room temperature for 12 hours and then filtered to recover a white solid which was washed with dichloromethane and dried to give the product as a white solid (2.35 g, Yield: 75%).

^{1 H NMR (300MHz, DMSO-} d 6): δ3.93 (s, 3H), 6.66 (t, J = 6.0Hz, 1H), 7.18 (d, J = 9.0Hz, 2H), 7.56 (d, J = 9.0 Hz, 2H), 7.59-7.63 (m, 2H), 7.98 (s, 1H), 8.10 (d, J = 1.8 Hz, 1H) , J = 1.8 Hz, 1 H), 8.98 (s, 1 H), 9.18 (s, 1 H)

MS (ESI +): m / z 505.8 [M + H] < ^{+ &}

Step 5: l- (4-Chloro-3- (trifluoromethyl) phenyl) -3- (4- (3-fluoro- -Yl-oxy) phenyl) urea Synthesis of p-toluenesulfonate:

To a solution of 1- (4-chloro-3- (trifluoromethyl) phenyl) -3- (4- (3-fluoro- (1.518 g, 3 mmol) and p-toluenesulfonic acid monohydrate (0.684 g, 3.6 mmol) was heated to reflux and anhydrous ethanol was added to dissolve the solid completely Respectively. The resulting clear solution was filtered and the filtrate was allowed to stand overnight, followed by suction filtration. The resulting white solid was collected and dried to give the product (1.328 g, yield: 65%) as a white solid.

^{1 H NMR (300MHz, DMSO-} d 6) δ9.32 (s, 1H), 9.12 (s, 1H), 8.39 (d, J = 1.2Hz, 1H), 8.28 (d, J = 5.7Hz, 1H) , 8.13 (d, J = 2.4 Hz, IH), 8.06 (s, IH), 7.72-7.56 (m, 4H), 7.51 (d, J = 8.0 Hz, 2H) 2H), 7.13 (d, J = 8.1 Hz, 2H), 6.77 (t, J = 6.2 Hz, 1H), 3.95 (s, 3H), 2.29 (s, 3H).

Example 2

FD-2013018:

3- (4- (3-Chloro-2- (l-methyl-pyrazol-4-yl) -pyridin- ) Phenyl) urea

Manufacturing method:

Step 1: Synthesis of 2,3-dichloro-4-iodopyridine

Butyllithium (2.4 M hexane solution, 59.12 mL, 141.9 mmol) was added dropwise to a solution of 2,3-dichloropyridine (20 g, 135.1 mmol) in anhydrous tetrahydrofuran (350 mL) at -78 캜 under an atmosphere of argon gas To obtain a reaction mixture, and the reaction mixture was stirred at -78 DEG C for 90 minutes. A solution of iodine (41 g, 161.5 mmol) in anhydrous tetrahydrofuran (100 mL) was added dropwise, and the reaction mixture was stirred at -78 째 C for 60 minutes. The temperature was then increased to room temperature. The reaction mixture was quenched with saturated ammonium chloride solution (100 mL), diluted with water (100 mL) and extracted with ethyl acetate (200 mL x 3). Combined organic phases. The combined organic phases were washed with brine (200 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 100: 1) to give the product (31.5 g, yield: 85.1%) as a yellow solid.

^{1 H NMR (300MHz, DMSO-} d 6): 8.08 (d, J = 5.4Hz, 1H), 8.65 (d, J = 5.4 Hz, 1H)

MS (ESI +): m / z 273.9 [M + H] < ^{+ &}

Step 2: Synthesis of 2,3-dichloro-4- (4-aminophenoxy) pyridine:

A solution of 4-aminophenol (13.85 g, 127.0 mmol) in anhydrous dimethylsulfoxide (120 mL) was bubbled with nitrogen gas for 10 minutes and then potassium tert-butoxide (13.60 g, 121.2 mmol) Respectively. The reaction mixture was stirred at room temperature for 30 minutes and then 2,3-dichloro-4-iodopyridine (31.5 g, 115.4 mmol) was added. The reaction mixture was stirred at room temperature for 5 hours, then diluted with water (500 mL) and extracted with ethyl acetate (300 mL x 3). Combined organic phases. The combined organic phases were washed with brine (300 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 4: 1, v / v) to give the product (27.5 g, yield: 93.7%) as a pale yellow solid.

^{1 H NMR (400 MHz, DMSO} -d 6): δ5.24 (br s, 2H), 6.64 (d, J = 8.8Hz, 2H), 6.67 (d, J = 5.6Hz, 1H), 6.90 (d , J = 8.8 Hz, 2H), 8.39 (d, J = 5.6 Hz, 1H)

MS (ESI +): m / z 255.0 [M + H] < ^{+ &}

Step 3: Synthesis of 4- (3-chloro-2- (1 -methyl-4-pyrazol-4-yl) -pyridin-

A solution of 2,3-dichloro-4- (4-aminophenoxy) pyridine (9.1 g, 35.7 mmol), 1-methylpyrazol-4-yl-boronic acid pinacol ester (7.42 mmol) in tetrahydrofuran (THF, A mixture of potassium carbonate (14.76 g, 106.9 mmol) and tetrakis (triphenylphosphine) palladium (2 g, 1.72 mmol) and water (35 mL) was bubbled with argon gas for 5 minutes, And the mixture was stirred under a gas atmosphere at 85 占 폚 for 24 hours to obtain a reaction mixture. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (100 mL x 3). Combined organic phases. The combined organic phases were washed with brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 1: 2, v / v) to give the product (4.85 g, yield: 45.2%) as a pale yellow solid.

^{1 H NMR (400MHz, DMSO-} d 6): δ3.93 (s, 3H), 5.19 (br s, 2H), 6.46 (d, J = 5.6Hz, 1H), 6.64 (d, J = 8.8Hz, 1H), 8.48 (s, 1H), 8.27 (d, J = 5.6 Hz,

MS (ESI +): m / z 301.0 [M + H] < ^{+ &}

Step 4: l- (4-Chloro-3- (trifluoromethyl) phenyl) -3- (4- Yl-oxy) phenyl) urea: < EMI ID =

To a solution of 4- (3-chloro-2- (l-methyl-pyrazol-4-yl) -pyridin-4-yl- oxy) aniline (4.85 g, 16.1 mmol) and 4- -Trifluoromethylphenyl isocyanate (4.28 g, 19.3 mmol) was stirred at room temperature for 12 hours and then filtered, and the resulting white solid was collected, washed with dichloromethane and dried to give the product as a white solid (7.2 g, : 85.5%).

^{1 H NMR (400MHz, DMSO-} d 6): δ3.94 (s, 3H), 6.56 (d, J = 5.2Hz, 1H), 7.19 (d, J = 8.8Hz, 2H), 7.59 (d, J 1H), 8.81 (s, 1H), 8.13 (s, 1H), 8.32 (d, J = 5.6Hz, 1H) (s, 1 H), 9.21 (s, 1 H)

MS (ESI +): m / z 522.1 [M + H] < ^{+ &}

Step 5: l- (4-Chloro-3- (trifluoromethyl) phenyl) -3- (4- Yl-oxy) phenyl) urea p-toluenesulfonate:

To a solution of l- (4-chloro-3- (trifluoromethyl) phenyl) -3- (4- (3-chloro- Phenyl) urea (1.570 g, 3 mmol) and p-toluenesulfonic acid monohydrate (0.684 g, 3.6 mmol) were heated to reflux and anhydrous ethanol was added until the solid was completely dissolved. The resulting clear solution was filtered, and the filtrate was left to stand overnight. The filtrate was then subjected to suction filtration, and the resulting white solid was recovered and dried to give the product (1.428 g, yield 69%) as a white solid.

1H NMR (300MHz, DMSO-d 6) δ9.33 (s, 1H), 9.14 (s, 1H), 8.56 (s, 1H), 8.42 - 8.32 (dd, J = 6.0, 2.4Hz, 1H), 8.15 (s, 1H), 8.13 (d, J = 2.4 Hz, 1H), 7.71-7.58 (m, 4H), 7.50 (d, J = 8.0 Hz, 2H) , 7.13 (d, J = 7.8 Hz, 2H), 6.71-6.60 (m, IH), 3.95 (s, 3H), 2.29 (s, 3H).

Example 3

FD-2013024:

3- (4- (3-Chloro-2- (l-methyl-pyrazol-4-yl) -pyridin- ) -2-fluorophenyl) urea

Manufacturing method:

Step 1: Synthesis of 2,3-dichloro-4- (4-amino-3-fluorophenoxy)

A solution of 4-amino-3-fluorophenol (1.69 g, 13.28 mmol) in anhydrous dimethyl sulfoxide (15 mL) was bubbled with nitrogen gas for 10 minutes, followed by the addition of 2,3-dichloro-4-iodopyridine , 12.13 mmol) was added to give a reaction mixture. The reaction mixture was stirred at room temperature for 30 minutes and then 2,3-dichloro-4-iodopyridine (31.5 g, 115.4 mmol) was added. The reaction mixture was stirred at room temperature for 5 hours, diluted with water (50 mL) and extracted with ethyl acetate (30 mL x 3). Combined organic phases. The combined organic phases were washed with brine (30 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 4: 1, v / v) to give the product (1.0 g, yield: 30%) as a pale yellow solid.

^{1 H NMR (300MHz, DMSO-} d 6) δ8.17 (d, J = 5.6Hz, 1H), 7.16-7.00 (m, 1H), 6.92-6.78 (m, 2H), 6.75 (d, J = 5.6 Hz, < / RTI > 1H), 5.26 (br s, 2H)

MS (ESI +): m / z 272.9 [M + H] < ^{+ &}

Step 2: Synthesis of 4- (3-chloro-2- (l -methyl-4-pyrazol-4-yl) -pyridin-4-yl-oxy) -2-fluoroaniline:

To a solution of 2,3-dichloro-4- (4-amino-3-fluorophenoxy) pyridine (0.40 g, 1.47 mmol), 1-methylpyrazol-4-yl-boronic acid (1 mL) and a mixture of potassium carbonate (0.70 g, 5.07 mmol) and tetrakis (triphenylphosphine) palladium (0.10 g, 0.086 mmol) Bubbled and stirred at 85 < 0 > C for 24 hours under an argon atmosphere to give a reaction mixture. The reaction mixture was diluted with water (200 mL) and extracted with ethyl acetate (20 mL x 3). Combined organic phases. The combined organic phases were washed with brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 1: 2, v / v) to give the product (0.23 g, yield: 49%) as a pale yellow solid.

^{1 H NMR (300MHz, DMSO-} d 6) δ8.48 (s, 1H), 8.29 (d, J = 5.5Hz, 1H), 8.11 (d, J = 0.6Hz, 1H), 7.04 (dd, J = 2H), 3.93 (s, 3H), 2.27 (d, J =

MS (ESI +): m / z 319.0 [M + H] < ^{+ &}

Step 3: l- (4-Chloro-3- (trifluoromethyl) phenyl) -3- (4- Yl-oxy) -2-fluorophenyl) urea: < EMI ID =

To a solution of 4- (3-chloro-2- (l-methyl-pyrazol-4-yl) -pyridin-4-yl-oxy) -2-fluoroaniline (0.23 g, 0.72 mmol) and 4-Chloro-3-trifluoromethylphenyl isocyanate (0.16 g, 0.72 mmol) was stirred at room temperature for 12 hours, then filtered and the resulting white solid was collected, washed with dichloromethane and dried to give the product as a white solid 0.30 g, yield: 77%).

^{1 H NMR (300MHz, DMSO-} d 6) δ9.52 (s, 1H), 8.75 (s, 1H), 8.52 (s, 1H), 8.36 (d, J = 5.5Hz, 1H), 8.26-8.05 ( 1H), 6.69 (d, J = 5.5 Hz, 1 H), 7.36 (dd, J = 11.5, 2.6 Hz, 3.94 (s, 3 H)

MS (ESI +): m / z 540.0 [M + H] < ^{+ &}

Example 4

FD-2013025:

3- (4- (3-Cyano-2- (l-methyl-pyrazol-4-yl) -pyridin- Oxy) phenyl) urea

Manufacturing method:

Step 1: Synthesis of 2-chloro-4-iodonicotinonitrile:

(2.4 M hexane solution, 3.0 mL, 7.2 mmol) was added dropwise to a solution of diisopropylamine (0.728 g, 7.2 mmol) in anhydrous tetrahydrofuran (20 mL) at -30 캜 under an argon atmosphere, &Lt; / RTI &gt; The reaction mixture was stirred at -30 占 폚 for 30 minutes, then cooled to -78 占 폚. A solution of 2-chloro-nicotinonitrile (1.0 g, 7.2 mmol) in anhydrous tetrahydrofuran (10 mL) was added dropwise and the reaction mixture was then stirred at -78 <0> C for 60 min. A solution of iodine (1.8 g, 7.2 mmol) in anhydrous tetrahydrofuran (10 mL) was added dropwise and the reaction mixture was then stirred at -78 <0> C for 30 min. The reaction mixture was quenched with saturated ammonium chloride solution (50 mL), diluted with water (50 mL) and extracted with ethyl acetate (100 mL x 3). Combined organic phases. The combined organic phases were washed with brine (100 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (petroleum ether: ethyl acetate = 80: 1) to give the product (0.357 g, yield: 19%) as a yellow solid.

^{1 H NMR (400MHz, DMSO-} d 6) δ8.32 (d, J = 5.2Hz, 1H), 8.15 (d, J = 5.2Hz, 1H)

MS (ESI +): m / z 264.9 [M + H] &lt; ^{+ &}

Step 2: Synthesis of 2-chloro-3-cyano-4- (4-aminophenoxy)

A solution of 4-aminophenol (164 mg, 1.48 mmol) in anhydrous dimethylsulfoxide (3 mL) was bubbled with nitrogen gas for 10 min followed by addition of potassium tert-butoxide (166 mg, 1.48 mmol) to give a reaction mixture. The reaction mixture was stirred at room temperature for 30 minutes and 2-chloro-4-iodonicotinonitrile (355 mg, 1.34 mmol) was added. The reaction mixture was stirred at room temperature for 5 hours, then diluted with water (30 mL) and extracted with ethyl acetate (30 mL x 3). Combined organic phases. The combined organic phases were washed with water (30 mL x 2), washed with brine (30 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 4: 1, v / v) to give the product (210 mg, yield: 61%) as a pale yellow solid.

^{1 H NMR (400MHz, DMSO-} d 6) δ8.42 (d, J = 6.0Hz, 1H), 6.96 (d, J = 8.8Hz, 2H), 6.76 (d, J = 6.0Hz, 1H), 6.65 (d, J = 8.8 Hz, 2H), 5.29 (s, 2H)

MS (ESI +): m / z 246.0 [M + H] &lt; ^{+ &}

Step 3: Synthesis of 4- (3-cyano- (1 -methyl-pyrazol-4-yl) -pyridin-

To a solution of 2-chloro-3-cyano-4- (4-aminophenoxy) pyridine (200 mg, 0.816 mmol), 1- methylpyrazol-4-yl-boronic acid pinacol ester A mixture of potassium carbonate (338 mg, 2.45 mmol) and tetrakis (triphenylphosphine) palladium (95 mg, 0.0816 mmol) and water (1 mL) was bubbled with argon gas for 5 minutes, And the mixture was stirred at 85 캜 for 24 hours under a gas atmosphere to obtain a reaction mixture. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL x 3). Combined organic phases. The combined organic phases were washed with water (20 mL x 2), washed with brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 1: 2, v / v) to give the product (95 mg, yield: 40%) as a pale yellow solid.

^{1 H NMR (300MHz, DMSO-} d 6) δ8.52 (d, J = 6.0Hz, 1H), 8.48 (s, 1H), 8.17 (s, 1H), 6.95 (d, J = 8.7Hz, 2H) , 6.65 (d, J = 8.7 Hz, 2H), 6.51 (d, J = 6.0 Hz,

MS (ESI +): m / z 292.1 [M + H] &lt; ^{+ &}

Step 4: 1- (4-Chloro-3- (trifluoromethyl) phenyl) -3- (4- (3-cyano- -Yl-oxy) phenyl) urea: &lt; EMI ID =

To a solution of 4- (3-cyano-2- (1-methyl-pyrazol-4-yl) -pyridin-4-yl- oxy) aniline (90 mg, 0.31 mmol) and 4- The resultant white solid was collected, washed with dichloromethane and dried to give the product (54 mg, yield: 34%) as a white solid. Trifluoromethylphenyl isocyanate (68.5 mg, 0.31 mmol) was stirred at room temperature for 12 hours, ).

^{1 H NMR (400MHz, DMSO-} d 6) δ9.23 (s, 1H), 9.05 (s, 1H), 8.56 (d, J = 6.0Hz, 1H), 8.51 (s, 1H), 8.19 (s, J = 9.0 Hz, 2H), 6.58 (d, J = 6.0 Hz, 1H), 3.96 (m, s, 3H)

MS (ESI +): m / z 512.9 [M + H] &lt; ^{+ &}

Example 5

FD-2013027:

(4-chloro-3- (trifluoromethyl) phenyl) -3- (4- (3- ) Phenyl) urea

Manufacturing method:

Step 1: Synthesis of 2-chloro-4-fluoro-3-methylpyridine:

Butyllithium (2.4 M hexane solution, 4.37 mL, 10.49 mmol) was added dropwise to a solution of diisopropylamine (1.06 g, 11 mmol) in anhydrous tetrahydrofuran (20 mL) at -30 캜 under an atmosphere of nitrogen gas, A mixture was obtained. The reaction mixture was stirred at -30 占 폚 for 30 minutes, then cooled to -78 占 폚. A solution of 2-chloro-4-fluoropyridine (1.31 g, 10 mmol) in anhydrous tetrahydrofuran (10 mL) was added dropwise, and then the reaction mixture was stirred at -78 ° C for 60 minutes. A solution of iodomethane (1.48 g, 10.5 mmol) in anhydrous tetrahydrofuran (5 mL) was added and the reaction mixture was then stirred at -78 <0> C for 30 min. The reaction mixture was quenched with saturated ammonium chloride solution (5 mL), diluted with water (50 mL) and extracted with ethyl acetate (30 mL x 3). Combined organic phases. The combined organic phases were washed with saturated brine (30 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (petroleum ether: ethyl acetate = 100: 1, v / v) to give the product (0.63 g, yield: 43%) as a yellow solid.

^{1 H NMR (300MHz, DMSO-} d 6) δ8.31 (dd, J = 8.0, 5.8Hz, 1H), 7.38 (dd, J = 8.7, 5.6Hz, 1H), 2.27 (d, J = 1.8Hz, 3H)

MS (ESI +): m / z 146.0 [M + H] &lt; ^{+ &}

Step 2: Synthesis of 2-chloro-3-methyl-4- (4-aminophenoxy)

A solution of 4-aminophenol (0.21 g, 1.91 mmol) in anhydrous dimethylsulfoxide (3 mL) was bubbled with nitrogen gas for 10 minutes and then potassium tert-butoxide (0.22 g, 1.96 mmol) Respectively. The reaction mixture was stirred at room temperature for 30 minutes and 2-chloro-4-fluoro-3-methylpyridine (269 mg, 1.85 mmol) was added. The reaction mixture was stirred at room temperature for 5 hours, then diluted with water (20 mL) and extracted with ethyl acetate (20 mL x 2). Combined organic phases. The combined organic phases were washed with water (20 mL x 2), washed with brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 5: 1, v / v) to give the product (0.38 g, yield: 88%) as a pale yellow solid.

^{1 H NMR (300MHz, DMSO-} d 6) δ8.04 (d, J = 5.6Hz, 1H), 6.89-6.78 (m, 2H), 6.69-6.57 (m, 2H), 6.51 (d, J = 5.7 Hz, &lt; / RTI &gt; 1H), 5.15 (s, 2H), 2.31 (s,

MS (ESI +): m / z 235.0 [M + H] &lt; ^{+ &}

Step 3: Synthesis of 4- (3-methyl-2- (1 -methyl-4-pyrazol-4-yl) -pyridin-

Methyl-4- (4-aminophenoxy) pyridine (380 mg, 1.62 mmol), 1-methyl-4-pyrazol- A mixture of the Cole ester (337 mg, 1.62 mmol), potassium carbonate (400 mg, 2.89 mmol) and tetrakis (triphenylphosphine) palladium (90 mg, 0.08 mmol) and water (1 mL) was bubbled with argon gas for 5 minutes, The reaction mixture was then stirred under an argon atmosphere at 85 DEG C for 24 hours. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL x 3). Combined organic phases. The combined organic phases were washed with water (20 mL x 2), washed with saturated brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 1: 2, v / v) to give the product (170 mg, yield: 37.5%) as a pale yellow solid.

^{1 H NMR (400MHz, DMSO-} d 6) δ8.18 (d, J = 5.6Hz, 1H), 8.17 (s, 1H), 7.88 (s, 1H), 6.82 (d, J = 8.7Hz, 2H) , 6.63 (d, J = 8.7 Hz, 2H), 6.35 (d, J = 5.6 Hz,

MS (ESI +): m / z 281.1 [M + H] &lt; ^{+ &}

Step 4: l- (4-Chloro-3- (trifluoromethyl) phenyl) -3- (4- Yl-oxy) phenyl) urea: &lt; EMI ID =

To a solution of 4- (3-methyl-2- (l-methyl-pyrazol-4-yl) -pyridin-4-yl- oxy) aniline (165 mg, 0.58 mmol) and 4- A solution of trifluoromethylphenyl isocyanate (155 mg, 0.7 mmol) was stirred at room temperature for 12 hours, then filtered and the white solid obtained was then recovered, washed with dichloromethane and dried to give the product (145 mg, yield: %).

^{1 H NMR (300MHz, DMSO-} d 6) δ9.18 (s, 1H), 8.94 (s, 1H), 8.24 (d, J = 5.4Hz, 1H), 8.19 (s, 1H), 8.12 (d, (D, J = 9.0 Hz, 2H), 7.09 (d, J = 9.0 Hz, 2H), 6.47 (d, J = 5.4 Hz, 1H), 3.92 (s, 3H), 2.40

MS (ESI +): m / z 501.9 [M + H] &lt; ^{+ &}

Example 6

FD-2013031:

(4-chloro-3- (trifluoromethyl) phenyl) -3- (4- (3- ) Phenyl) urea

Manufacturing method:

Step 1: Synthesis of 2-chloro-3-nitro-4- (4-aminophenoxy)

A solution of 4-aminophenol (1.09 g, 10 mmol) in anhydrous dimethylsulfoxide (10 mL) was bubbled with nitrogen gas for 10 min followed by addition of potassium tert-butoxide (1.12 g, 10 mmol) to give a reaction mixture. The reaction mixture was stirred at room temperature for 15 minutes and 2,4-dichloro-3-nitropyridine (1.93 g, 10 mmol) was added. The reaction mixture was stirred at room temperature for 5 hours, then diluted with water (100 mL) and extracted with ethyl acetate (50 mL x 3). Combined organic phases. The combined organic phases were washed with water (50 mL x 2), washed with brine (50 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 3: 1, v / v) to give the product (408 mg, yield: 15%) as a pale yellow solid.

^{1 H NMR (300MHz, DMSO-} d 6) δ8.40 (d, J = 5.7Hz, 1H), 6.33 (d, J = 8.7Hz, 2H), 6.28 (d, J = 5.7Hz, 2H), 6.63 (d, J = 8.7 Hz, 2H), 5.30 (br s, 2H)

MS (ESI +): m / z 266.0 [M + H] &lt; ^{+ &}

Step 2: Synthesis of 4- (3-nitro-2- (1-methyl-pyrazol-4-yl) -pyridin-

To a solution of 2-chloro-3-nitro-4- (4-aminophenoxy) pyridine (1.25 mg, 1.51 mmol), 1- methyl- pyrazol- A mixture of ester (377 mg, 1.81 mmol), potassium carbonate (12.4 g, 9.0 mmol) and tetrakis (triphenylphosphine) palladium (174 mg, 1.151 mmol) and water (3 mL) was bubbled with argon gas for 5 minutes, The reaction mixture was stirred overnight at 85 캜 under an argon atmosphere. The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (30 mL x 3). Combined organic phases. The combined organic phases were washed with water (30 mL x 2), washed with saturated brine (30 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 1: 3, v / v) to give the product (450 mg, yield: 96%) as a pale yellow solid.

^{1 H NMR (300MHz, DMSO-} d 6) δ8.49 (d, J = 5.7Hz, 1H), 8.10 (s, 1H), 7.72 (s, 1H), 7.69-7.46 (m, 11H), 6.91 ( (d, J = 8.7 Hz, 2H), 6.68 (d, J = 5.7 Hz, 1H), 6.63

MS (ESI +) m / z 312.0 [M + H] &lt; ^{+ &}

Step 3: Synthesis of 4- (3-amino-2- (1-methyl-pyrazol-4-yl) -pyridin-

(200 mg, 0.64 mmol) and palladium-carbon (20 mg) in anhydrous methanol (15 mL) was added to a solution of 4- (3-nitro- Was stirred at room temperature under an atmosphere of 4 atm for 4 hours. The palladium-carbon was filtered through Celite and then the filtrate was concentrated. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 1: 4) to give the product (75 mg, yield: 41%).

^{1 H NMR (400MHz, DMSO-} d 6) δ8.20 (s, 1H), 7.91 (s, 1H), 7.73 (d, J = 5.2Hz, 1H), 6.83 (d, J = 8.8Hz, 2H) 2H), 6.62 (d, J = 8.8 Hz, 2H), 6.29 (d, J = 5.2 Hz,

MS (ESI +): m / z 282.1 [M + H] &lt; ^{+ &}

Step 4: l- (4-Chloro-3- (trifluoromethyl) phenyl) -3- (4- Yl-oxy) phenyl) urea: &lt; EMI ID =

To a solution of 4- (3-amino-2- (l-methyl-pyrazol-4-yl) -pyridin-4-yl- oxy) aniline (70 mg, 0.249 mmol) and 4- The resulting white solid was collected, washed with dichloromethane and dried to give the product (79 mg, yield: 63%) as a white solid. The title compound was obtained as a white solid &Lt; / RTI &gt;

^{1 H NMR (300MHz, DMSO-} d 6) δ9.17 (s, 1H), 8.92 (s, 1H), 8.23 (s, 1H), 8.11 (s, 1H), 7.93 (s, 1H), 7.78 ( J = 5.4 Hz, 1H), 7.63 (d, J = 2.6 Hz, 2H), 7.67-7.59 (m, 2H), 7.10 Hz, 1 H), 4.82 (br s, 2 H), 3.91 (s, 3 H)

MS (ESI +): m / z 502.9 [M + H] &lt; ^{+ &}

Example 7

FD-2013033:

3- (4- (3-methylamino-2- (l-methyl-4-pyrazol-4-yl) -pyridin- Yl-oxy) phenyl) urea

Manufacturing method:

Step 1: Synthesis of 2-chloro-4-iodo-3- (methylamino) pyridine:

A solution of 2-chloro-3-fluoro-4-iodopyridine (12 g, 46.6 mmol) in aminomethane and ethanol (25%, v / v, 30 mL) was stirred at 65 ° C for 8 hours. The volatiles were removed under reduced pressure. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 30: 1) to give the product (5.5 g, yield: 44%) as yellow oil.

^{1 H NMR (300MHz, DMSO-} d 6) δ7.77 (d, J = 4.9Hz, 1H), 7.51 (d, J = 4.9Hz, 1H), 4.75 (br, s, 1H), 2.91 (s, 3H)

MS (ESI +): m / z 268.9 [M + H] &lt; ^{+ &}

Step 2: Synthesis of 2-chloro-3-methylamino-4- (4-aminophenoxy)

4-Aminophenol (0.16 g, 1.46 mmol) in anhydrous dimethylsulfoxide (3 mL) was bubbled with nitrogen gas for 10 min and then potassium tert-butoxide (0.16 g, 1.46 mmol) was added to give a reaction mixture . The reaction mixture was stirred at room temperature for 30 minutes and then 2-chloro-4-iodo-3- (methylamino) pyridine (170 mg, 0.63 mmol) was added. The reaction mixture was stirred at room temperature for 1 hour and then at 80 ° C for 5 hours. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL x 3). Combined organic phases. The combined organic phases were washed with water (20 mL x 2), washed with brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate 5: 1, v / v) to give the product (96 mg, yield: 61%) as a pale yellow solid.

^{1 H NMR (400MHz, DMSO-} d 6) δ7.57 (d, J = 5.3Hz, 1H), 6.81 (d, J = 8.8Hz, 2H), 6.66-6.56 (m, 2H), 6.48 (d, J = 5.3 Hz, 1H), 5.08 (s, 2H), 4.94 (q, J = 5.4 Hz,

MS (ESI +): m / z 250.0 [M + H] &lt; ^{+ &}

Step 3: Synthesis of 4- (3-methylamino-2- (1-methyl-pyrazol-4-yl) -pyridin-

To a solution of 2-chloro-3-methylamino-4- (4-aminophenoxy) pyridine (270 mg, 1.08 mmol), 1-methylpyrazol-4-yl-boronic acid pinacol ester (1 mL) and a mixture of potassium carbonate (447 mg, 3.24 mmol) and bis (triphenylphosphine) dichloropalladium (II) (76 mg, 0.108 mmol) were bubbled with argon gas for 5 minutes , Followed by stirring at 85 캜 for 24 hours under an atmosphere of argon gas to obtain a reaction mixture. The reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (30 mL x 3). Combined organic phases. The combined organic phases were washed with water (30 mL x 2), washed with saturated brine (30 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 1: 3, v / v) to give the product (180 mg, yield: 56%) as a pale yellow solid.

^{1 H NMR (400MHz, DMSO-} d 6) δ8.19 (s, 1H), 8.02 (s, 1H), 7.94 (s, 1H), 7.91 (d, J = 5.4Hz, 1H), 6.84 (d, 2H), 6.62 (d, J = 8.7 Hz, 2H), 6.35 (d, J = 5.4 Hz, 1H), 5.14 (s, 3H)

MS (ESI +): m / z 296.1 [M + H] &lt; ^{+ &}

Step 4: l- (4-Chloro-3- (trifluoromethyl) phenyl) -3- (4- (3- methylamino- -Yl-oxy) phenyl) urea: &lt; EMI ID =

To a solution of 4- (3-methylamino-2- (l-methyl-pyrazol-4-yl) -pyridin-4-yl- oxy) aniline (170 mg, 0.576 mmol) and 4- -Trifluoromethylphenyl isocyanate (127.6 mg, 0.576 mmol) was stirred at room temperature for 12 hours and then filtered. The resulting white solid was collected, washed with dichloromethane and dried to give the product (67 mg, yield: 22.5%).

^{1 H NMR (300MHz, DMSO-} d 6) δ9.16 (s, 1H), 8.91 (s, 1H), 8.20 (s, 1H), 8.11 (s, 1H), 7.95 (d, J = 5.1Hz, J = 9.0 Hz, 2H), 7.94 (s, 1H), 7.66-7.60 (m, 2H), 7.53 (d, 3H), 2.69 (d, J = 4.8 Hz, 3H), 3.91 (s,

MS (ESI +): m / z 517.1 [M + H] &lt; ^{+ &}

Example 8

FD-2013037:

3- (4- (3-Methoxy-2- (l-methyl-pyrazol-4-yl) -pyridin- Oxy) phenyl) urea

Manufacturing method:

Step 1: Synthesis of 2-chloro-3-methoxy-4-iodopyridine:

2-Chloro-3-fluoro-4-iodopyridine (1.05 g, 4.08 mmol) and sodium methoxide (0.22 g, 4.08 mmol) in methanol (10 mL) were stirred at 45 ° C for 2 hours to give a reaction mixture . The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL x 3). Combined organic phases. The combined organic phases were washed with water (50 mL x 2), washed with brine (50 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 100: 1, v / v) to give the product (0.58 g, yield: 53%) as a pale yellow solid.

^{1 H NMR (300MHz, DMSO-} d 6) δ7.92 (d, J = 5.1Hz, 1H), 7.86 (d, J = 5.1Hz, 1H), 3.83 (s, 3H)

MS (ESI +): m / z 269.8 [M + H] &lt; ^{+ &}

Step 2: Synthesis of 2-chloro-3-methoxy-4- (4-aminophenoxy)

(0.172 g, 1.58 mmol), 2-chloro-3-methoxy-4-iodopyridine (425 mg, 1.58 mmol) and potassium tert-butoxide in anhydrous dimethylsulfoxide (0.177 g, 1.58 mmol) was stirred at 155 占 폚 for 5 hours to obtain a reaction mixture. The reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (30 mL x 3). Combined organic phases. The combined organic phases were washed with water (30 mL x 2), washed with brine (30 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate 4: 1, v / v) to give the product (210 mg, yield 53%) as a light yellow solid.

¹ H NMR (300MHz, DMSO-d ₆₎ δ7.95 (d, J = 5.7Hz, 1H), 6.91- 6.86 (d, J = 8.7Hz, 2H), 6.63 (d, = J 8.7Hz, 2H) , 6.62 (d, J = 5.7 Hz, 1 H), 5.16 (s, 2H), 3.90

MS (ESI +): m / z 251.0 [M + H] &lt; ^{+ &}

Step 3: Synthesis of 4- (3-methoxy-2- (1-methyl-pyrazol-4-yl) -pyridin-

To a solution of 2-chloro-3-methoxy-4- (4-aminophenoxy) pyridine (130 mg, 0.52 mmol), 1-methylpyrazol-4-yl-boronic acid pinacol ester (1 mL) and a mixture of potassium carbonate (120 mg, 0.58 mmol), potassium carbonate (215 mg, 1.56 mmol) and bis (triphenylphosphine) dichloropalladium (II) (90 mg, 0.127 mmol) were bubbled with argon gas for 5 minutes , Followed by stirring at 100 DEG C for 5 minutes in an atmosphere of argon gas to obtain a reaction mixture. The reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (30 mL x 3). Combined organic phases. The combined organic phases were washed with water (30 mL x 2), washed with brine (30 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate 1: 3, v / v) to give the product (40 mg, yield: 26%) as a pale yellow solid.

^{1 H NMR (300MHz, DMSO-} d 6) δ8.27 (s, 1H), 8.07 (d, J = 5.4Hz, 1H), 8.00 (s, 1H), 6.87 (d, J = 8.7Hz, 2H) , 6.62 (d, J = 8.7 Hz, 2H), 6.41 (d, J = 5.4 Hz, 1H)

MS (ESI +): m / z 297.1 [M + H] +

Step 4: l- (4-Chloro-3- (trifluoromethyl) phenyl) -3- (4- (3-methoxy- -Yl-oxy) phenyl) urea: &lt; EMI ID =

To a solution of 4- (3-methoxy-2- (l-methyl-pyrazol-4-yl) -pyridin-4-yl- oxy) aniline (40 mg, 0.135 mmol) and 4- -Trifluoromethylphenyl isocyanate (30 mg, 0.135 mmol) in dichloromethane (10 ml) was stirred at room temperature for 12 hours, then filtered and the resulting white solid was recovered, washed with dichloromethane and dried to give the product (50 mg, 71%).

^{1 H NMR (300MHz, DMSO-} d 6) δ9.18 (s, 1H), 8.95 (s, 1H), 8.30 (s, 1H), 8.17-8.08 (m, 2H), 8.02 (s, 1H), J = 9.0 Hz, 2H), 7.55 (d, J = 9.0 Hz, 2H), 7.15 ), 3.89 (s, 3H)

MS (ESI +): m / z 517.9 [M + H] &lt; ^{+ &}

Comparative Example 1

Comparative compound: Soraphenylb free base prepared by the method as disclosed in the patent application document WO0042012A1

Comparative Example 2

Compound No. FD-1210005

The compound of Example 1 as disclosed in CN201110435847.9

Manufacturing method:

Step 1: Synthesis of 2-chloro-4- (4-aminophenoxy) pyridine:

A solution of 4-aminophenol (4.35 g, 39.8 mmol) in 40 mL of anhydrous dimethyl sulfoxide was bubbled with nitrogen gas for 10 minutes, followed by potassium tert-butoxide (4.7 g, 41.8 mmol) And then 2-chloro-4-fluoropyridine (5 g, 38.0 mmol) was added to give a reaction mixture. The reaction mixture was slowly heated at 80 &lt; 0 &gt; C and reacted at that temperature for 2 hours, cooled to room temperature when TLC showed the reaction was complete, then diluted with water (100 mL) and extracted with ethyl acetate (100 mL x 3) . The ethyl acetate phases were combined, washed with water (100 mL x 2), washed again with brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate 4: 1, v / v) to give the product (7.26 g, yield 86.8%) as a pale yellow solid.

_{^{1 H NMR (300MHz, CDCl 3}} ): δ4.07 (br s, 2H), 6.72 (d, J = 8.7Hz, 2H), 6.75-6.77 (m, 2H), 6.88 (d, J = 8.7Hz, 2H), 8.19 (d, J = 5.4 Hz, 1 H).

MS (ESI +): 221.1 [M + H] &lt; ^{+ &}

Step 2: Synthesis of 4- (2- (1-methyl-4-pyrazol-4-yl) -pyridin-

(6.47 g, 31.1 mmol) and 2-chloro-4- (4-aminophenoxy) pyridine (5.7 g, 25.9 mmol) Was dissolved in tetrahydrofuran (THF, 70 mL). Under stirring, potassium carbonate (10.7 g, 77.5 mmol) and water (17.1 mL) were added followed by tetrakis (triphenylphosphine) palladium (1.5 g, 1.29 mmol) , Stirred for 24 hours, cooled to room temperature, concentrated and then diluted with water (50 mL) and extracted with ethyl acetate (50 mL x 3). The ethyl acetate phases were combined, washed with water (50 mL x 2), washed again with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate 1: 2, v / v) to give the product (5.85 g, yield: 85%) as a pale yellow solid.

¹ H NMR (300MHz, CDCl _3): δ3.84 (s br, 2H), 3.92 (s, 3H), 6.60 (dd, = J 2.4, 5.7Hz, 1H), 6.71 (d, = J 8.7Hz, 2H), 6.91 (d, J = 8.7 Hz, 2H), 6.94 (d, J = 2.1 Hz, 1H).

MS (ESI &lt; ⁺ &gt;): 267.1 [M + H]

Step 3: l- (4-Chloro-3- (trifluoromethyl) phenyl) -3- (4- Phenyl) &lt; / RTI &gt; urea:

(6.7 g, 25.1 mmol) was dissolved in ethyl acetate (80 mL) and, under the protection of nitrogen gas, a solution of 4- (2- (1- methyl- 3-Trifluoromethyl-4-chloro-phenyl isocyanate (5.6 g, 25.1 mmol) was added, and after stirring at room temperature for 12 hours, a large amount of solid was precipitated. The reaction mixture was concentrated until the volume of the solvent was 40 mL, suction filtered, washed with ethyl acetate and dried to give the product (7.5 g, yield: 60.9%) as a white solid.

^{1 H NMR (300MHz, DMSO-} d 6): δ3.88 (s, 3H), 6.63 (d, J = 3.9Hz, 1H), 7.15 (d, J = 8.4Hz, 2H), 7.21 (s, 1H ), 7.57-7.69 (m, 4H), 7.96 (s, IH), 8.12 (s, IH), 8.24 (s, IH), 8.37 (d, J = 5.4 Hz, , 9.17 (s, 1 H).

MS (ESI &lt; ⁺ &gt;): 488.1 [M + H]

Comparative Example 3

Compound No. FD-2013016.

3- (4- (2- (1 -methyl-pyrazol-4-yl) -5-fluoro-pyridin- Oxy) phenyl) urea;

Manufacturing method:

Step 1: Synthesis of 2-chloro-4-iodo-5-fluoropyridine

In a 100 mL three necked flask, 2-chloro-5-fluoropyridine (2.65 g, 20.1 mmol) was dissolved in anhydrous tetrahydrofuran (30 mL) and protected under nitrogen gas at- And stirred for 30 minutes. A solution of tert-butyllithium in n-pentane (16.27 mL, 21.1 mmol) was slowly added dropwise, and after the addition, the reaction was allowed to react at that temperature for 90 minutes. Subsequently, a solution of iodine (6.13 g, 24.2 mmol) in anhydrous tetrahydrofuran (10 mL) was slowly added dropwise. After the addition, the temperature was gradually increased to room temperature. Saturated ammonium chloride solution (100 mL) was added followed by water (50 mL), followed by phase separation and the water phase was extracted once with 50 mL, 40 mL, and 200 mL of ethyl acetate separately . The organic phases were combined, and the organic phase was washed with a saturated sodium thiosulfate solution (50 mL x 2), washed with brine (50 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 100: 1, v / v) to give the product (2 g, yield 39%) as a yellow solid.

MS (ESI &lt; ⁺ &gt;): 257.9 [M + H]

Step 2: Synthesis of 2-chloro-5-fluoro-4- (4-aminophenoxy)

4-Aminophenol (0.55 g, 5 mmol) was dissolved in anhydrous dimethylsulfoxide (15 mL), purged with nitrogen gas for 10 min and then potassium t-butoxide (0.58 g, 5.2 mmol) After stirring at room temperature for 30 minutes, 2-chloro-4-iodo-5-fluoropyridine (1.3 g, 5 mmol) was added and the reaction was allowed to proceed at room temperature for 5 hours. TLC showed the reaction was complete. Ethyl acetate (50 mL) was added, followed by sufficient stirring, followed by addition of water (100 mL), separation was performed, and the water phase was extracted with ethyl acetate (50 mL x 3). The ethyl acetate phases were combined, washed with water (50 mL x 2), washed again with brine (50 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate 4: 1, v / v) to give the product (0.2g yield: 16.8%) as a pale yellow solid.

^{1 H NMR (300MHz, DMSO-} d 6): δ5.24 (br s, 2H), 6.64 (d, J = 8.7Hz, 2H), 6.65 (s, 1H), 6.94 (d, J = 8.7Hz, 2H), 8.44 (d, J = 3.0 Hz, 1H).

MS (ESI &lt; ⁺ &gt;): 239.1 [M + H]

Step 3: Synthesis of 4- (5-fluoro-2- (1-methyl-pyrazol-4-yl) -pyridin-

To a solution of 2-chloro-5-fluoro-4- (4-aminophenoxy) pyridine (200 mg, 0.84 mmol) and 1-methylpyrazole-4- boronic acid pinacol ester (175 mg, 0.84 mmol ) Was dissolved in tetrahydrofuran (THF, 5 mL), potassium carbonate (347 mg, 2.51 mmol) and water (0.84 mL) were added and oxygen was removed. Under the protection of argon gas, tetrakis Phosphine) palladium (48 mg, 0.04 mmol) was added in the dark and stirred at 85 &lt; 0 &gt; C for 24 h, then cooled to room temperature when TLC showed the reaction was complete. Ethyl acetate and water were then added (20 mL each), phase separation was performed, and the aqueous phase was extracted with ethyl acetate (20 mL x 2). The ethyl acetate phases were combined, washed with brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate 4: 1, v / v) to give the product (50 mg, yield: 21%) as a light yellow solid.

^{1 H NMR (300 MHz, DMSO} -d 6): δ3.82 (s, 3H), 5.16 (br s, 2H), 6.63 (d, J = 8.7Hz, 2H), 6.89 (d, J = 6.6Hz , 6.91 (d, J = 8.7 Hz, 2H), 7.76 (s, 1H), 8.10 (s, 1H), 8.46 (d, J = 3.0 Hz, 1H).

MS (ESI +): 285.0 [M + H] &lt; ⁺ &gt;

Step 4: l- (4-Chloro-3- (trifluoromethyl) phenyl) -3- (4- (5- fluoro- -Yl-oxy) phenyl) urea: &lt; EMI ID =

(50 mg, 0.176 mmol) was dissolved in dichloromethane (2 mL), and a solution of 4- (5-fluoro-2- (1- methyl-pyrazol- Was added 4-chloro-3-trifluoromethylphenyl isocyanate (46 mg, 0.208 mmol) under stirring and the mixture was stirred at room temperature for 12 hours. A large amount of solid was precipitated, filtered off with suction, washed with dichloromethane and dried to give the product (61 mg, yield: 68.6%) as a white solid.

^{1 HNMR (300MHz, DMSO-d} 6): δ3.83 (s, 3H), 7.11 (d, J = 6.6Hz, 1H), 7.18 (d, J = 9.0Hz, 2H), 7.56 (d, J = 2H), 7.60-7.67 (m, 2H), 7.83 (s, 1H), 8.11 (d, J = 2.1 Hz, 1H) 1H), 8.95 (s, 1 H), 9.17 (s, 1 H).

MS (ESI &lt; ⁺ &gt;): 506.1 [M + H]

Step 5: l- (4-Chloro-3- (trifluoromethyl) phenyl) -3- (4- (5-fluoro- -Yl-oxy) phenyl) urea Synthesis of p-toluenesulfonate:

4- (5-Fluoro-2- (l-methyl-pyrazol-4-yl) -pyridin- Oxy) phenyl) urea (55 mg, 0.109 mmol) and p-toluenesulfonic acid monohydrate (25 mg, 0.131 mmol) were added to anhydrous ethanol (2 mL). The resulting mixture was heated to reflux and anhydrous ethanol was added until the solid was completely dissolved. The resulting clear solution was filtered and the filtrate was allowed to stand overnight, followed by suction filtration. The resulting white solid was recovered and dried to give the product (42 mg, yield: 57%) as a white solid.

1H), 8.23 (s, 1H), 8.12 (d, J = 1.7 Hz, 1H) , 7.90 (s, 1H), 7.74-7.55 (m, 4H), 7.50 (dd, J = 5.3, 2.8Hz, 2H), 7.26-7.07 2.29 (s, 1 H).

Comparative Example 4

Compound No. FD-2013019.

4- (2-methyl-4-pyrazol-4-yl) -5-chloro-pyridin- -Oxy) phenyl) urea;

Manufacturing method:

Step 1: Synthesis of 2,5-dichloro-4-iodopyridine

In a 100 mL three-necked flask, 2,5-dichloropyridine (3.0 g, 20.3 mmol) was dissolved in anhydrous tetrahydrofuran (30 mL) and the temperature was reduced to -78 ° C under the protection of nitrogen gas. After 30 minutes, a 2.4 M n-butyllithium solution in n-hexane (8.9 mL, 21.3 mmol) was slowly added dropwise. After the addition, the reaction was carried out at that temperature for 90 minutes and then the iodine solution (6.13 g, 24.2 mmol) in anhydrous tetrahydrofuran (10 mL) was slowly added dropwise to the reaction system and the temperature was then increased to room temperature. Saturated ammonium chloride solution (100 mL) was added and water (50 mL) was also added, followed by phase separation and the aqueous phase was extracted once with 50 mL, 40 mL, and 200 mL of ethyl acetate, respectively. The combined organic phases were washed with saturated sodium thiosulfate solution (50 mL x 2) and brine (50 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product (4 g, yield: 72 %). The crude product was used directly in the next step without further purification.

MS (ESI &lt; ⁺ &gt;): 273.9 [M + H]

Step 2: Synthesis of 2,5-dichloro-4- (4-aminophenoxy) pyridine:

4-Aminophenol (1.59 g, 14.6 mmol) was dissolved in anhydrous dimethylsulfoxide (30 mL), purged with nitrogen gas for 10 minutes, potassium t-butoxide (1.68 g, 15 mmol) Stirred, and 2,5-dichloro-4-iodopyridine (4 g, 14.6 mmol) was added, followed by reaction at room temperature for 5 hours. TLC showed complete reaction. Ethyl acetate (80 mL) was added and stirred well, followed by water (100 mL). After phase separation, the aqueous phase was extracted with ethyl acetate (100 mL x 3).

The ethyl acetate phases were combined, washed with water (150 mL x 2), washed with brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate 4: 1, v / v) to give the product (0.34 g, yield: 9.1%) as a pale yellow solid.

MS (ESI +): 255.0 [M + H] &lt; ⁺ &gt;

Step 3: Synthesis of 4- (5-chloro-2- (l-methyl-pyrazol-4-yl) -pyridin-

(340 mg, 1.33 mmol) and 1-methylpyrazol-4-yl-boronic acid pinacol ester (278 mg, 1.33 mmol) were added under the protection of nitrogen gas. Was dissolved in tetrahydrofuran (THF, 8 mL), potassium carbonate (548 mg, 3.97 mmol) and water (1.33 mL) were added followed by tetrakis (triphenylphosphine) palladium (76 mg, 0.06 mmol) Was added in the dark and then stirred at 85 [deg.] C for 24 hours and when TLC showed the reaction was complete, it was cooled to room temperature and ethyl acetate and water were added (20 mL for each). After phase separation, the aqueous phase was extracted with ethyl acetate (20 mL x 2). The ethyl acetate phases were combined, washed with brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate 4: 1, v / v) to give the product (75 mg, yield: 19%) as a pale yellow solid.

MS (ESI +): 301.0 [M + H] &lt; ^{+ &}

Step 4: l- (4-Chloro-3- (trifluoromethyl) phenyl) -3- (4- Yl-oxy) phenyl) urea: &lt; EMI ID =

(75 mg, 0.25 mmol) was dissolved in dichloromethane (2 mL), and a solution of 4- (5-chloro-2- (1- methyl-pyrazol-4-yl) -pyridin- Under protection, 4-chloro-3-trifluoromethylphenyl isocyanate (67 mg, 0.3 mmol) was added, followed by stirring at room temperature for 12 hours, followed by precipitation of a large amount of solid, followed by suction filtration, washing with dichloromethane , And dried to give the product (78 mg, yield: 59.7%) as a white solid.

^{1 H NMR (300MHz, DMSO-} d 6): δ3.82 (s, 3H), 6.98 (s, 1H), 7.18 (d, J = 9.0Hz, 2H), 7.59 (d, J = 9.0Hz, 2H 1H, s), 7.63-7.66 (m, 2H), 7.82 (s, 1H), 8.12 (d, J = 2.1 Hz, 1H) , 9.25 (s, 1 H).

MS (ESI &lt; ⁺ &gt;): 522.1 [M + H]

Step 5: l- (4-Chloro-3- (trifluoromethyl) phenyl) -3- (4- (5- -Oxy) phenyl) urea Synthesis of p-toluenesulfonate:

(1-methyl-4-pyrazol-4-yl) -pyridin-4-yl -Oxy) phenyl) urea (70 mg, 0.134 mmol) and p-toluenesulfonic acid monohydrate (31 mg, 0.161 mmol) were added to anhydrous ethanol (2 mL). The resulting mixture was heated to reflux and anhydrous ethanol was further added until the solid was completely dissolved. The resulting clear solution was filtered, and the filtrate was left standing overnight. The filtrate was then suction filtered, and the resulting white solid was recovered and dried to give the product (57 mg, yield: 61%) as a white solid.

^{1 H NMR (300MHz, DMSO)} δ9.25 (s, 1H), 9.05 (s, 1H), 8.59 (s, 1H), 8.22 (s, 1H), 8.12 (d, J = 2.2Hz, 1H), J = 8.0 Hz, 2H), 7.18 (d, J = 9.0 Hz, 2H), 7.11 (d, J = 7.9 Hz, 2H), 7.87 (s, 1H), 7.72-7.54 ), 6.98 (s, 1 H), 3.82 (s, 3 H), 2.29 (s, 3 H).

Comparative Example 5

Compound No. FD-2013017

The title compound was prepared from 1- (4-chloro-3- (trifluoromethyl) phenyl) -3- (4- (2- ) Phenyl) urea

Manufacturing method:

Step 1: 2,6-Dichloro-4- (4-aminophenyl) pyridine

4-Aminophenol (2.39 g, 21.9 mmol) was dissolved in anhydrous dimethyl sulfoxide (30 mL), purged with nitrogen gas for 10 minutes, potassium t-butoxide (2.45 g, 21.9 mmol) was added, And stirred for 30 minutes. 2,4,6-Trichloropyridine (4 g, 21.9 mmol) was added, followed by reaction at 45 ° C for 5 hours, indicating that the reaction was complete by TLC. Ethyl acetate (80 mL) was added and stirred well, followed by addition of water (100 mL) followed by phase separation. The aqueous phase was extracted with ethyl acetate (100 mL x 3). The ethyl acetate phases were combined and washed with water (150 mL x 2), washed with brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product (5.1 g, yield: 91% ). The crude product was used directly in the next step without further purification.

MS (ESI +): 255.0 [M + H] &lt; ⁺ &gt;

Step 2: Synthesis of 2-chloro-6-methoxy-4- (4-aminophenoxy)

Dissolve 2,6-dichloro-4- (4-aminophenyl) pyridine (7.2 g, 28.2 mmol) in anhydrous methanol (50 mL), add sodium methoxide (1.52 g, 28.2 mmol) , Followed by distillation and drying under reduced pressure. Water (100 mL) was then added. The obtained solution was extracted with ethyl acetate (100 mL x 3), washed with brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (petroleum ether: ethyl acetate 8: 1) to give the product (0.88 g, yield: 12%) as a yellow solid.

MS (ESI &lt; ⁺ &gt;): 251.0 [M + H]

Step 3: Synthesis of 4- (6-methoxy-2- (1-methyl-pyrazol-4-yl) -pyridin-

(440 mg, 1.76 mmol) and 1-methylpyrazol-4-yl-boronic acid pinacol ester (368 mg, 1.76 mmol) were added under the protection of nitrogen gas to a solution of 2-chloro-6-methoxy- 1.76 mmol) was dissolved in tetrahydrofuran (THF, 8 mL), potassium carbonate (726 mg, 5.25 mmol) and water (1.76 mL) were added followed by tetrakis (triphenylphosphine) palladium 0.08 mmol) was added in the dark and after stirring at 85 [deg.] C for 24 h, TLC was cooled to room temperature when the reaction was complete. Then, ethyl acetate and water (20 mL each) were added followed by phase separation. The aqueous phase was extracted with ethyl acetate (20 mL x 2), and the ethyl acetate phases were combined, washed with brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate 4: 1, v / v) to give the product (400 mg, yield: 77%) as a pale yellow solid.

MS (ESI &lt; ⁺ &gt;): 297.2 [M + H]

Step 4: l- (4-Chloro-3- (trifluoromethyl) phenyl) -3- (4- (6-methoxy- -Yl-oxy) phenyl) urea: &lt; EMI ID =

(400 mg, 1.35 mmol) was dissolved in dichloromethane (2 mL), and a solution of 4- (6-methoxy-2- 4-Chloro-3-trifluoromethylphenyl isocyanate (300 mg, 1.35 mmol) was added under the protection of a gas and stirred at room temperature for 12 hours. Subsequently, the solid was precipitated, suction filtered, washed with dichloromethane, dried To give the product (300 mg, yield: 43%) as a white solid.

^{1 H NMR (300MHz, DMSO-} d 6): δ3.84 (s, 3H), 3.85 (s, 3H), 6.25 (d, J = 2.4Hz, 1H), 7.00-7.03 (m, 1H), 7.12 (d, J = 8.4 Hz, 2H), 7.51 (d, J = 8.4 Hz, 2H), 7.60-7.68 (m, 2H), 7.86 J = 2.4 Hz, 1 H), 8.94 (s, 1 H), 9.23 (s, 1 H).

MS (ESI &lt; ⁺ &gt;): 518.1 [M + H]

Experimental Example 1: Determination of compound for inhibitory activity of VEGFR2 kinase

1. Materials and equipment

EnVision 2104 multi-label microplate detector (PerkinElmer);

OptiPlate-384 White Opaque 384-well microwell plate (Cat. 6007290, PerkinElmer);

HTRF®KinEASE ^™ -TK kit (Cat.62 TKOPEC, Cisbio);

VEGFR2 (Cat: k2643, Sigma);

5x Kinase buffer (Cat: PV3189, Invitrogen);

ATP 10mM (Cat.PV3227, Invitrogen);

DTT 1M (Cat.D5545, Sigma);

MgCl ₂ 1M (Cat.M8266, Sigma);

MnCl ₂ 1M (Cat. 244589, Sigma);

Test compounds: The compounds prepared in the examples

Control compounds: The compounds prepared in the comparative examples

2. Experimental steps

2.1 Preparation of VEGFR2 kinase reagent

Table 1 Components and their concentrations in the VEGFR2 kinase reaction system

1 × Kinase buffer: 1 ml of 1 × Kinase buffer containing ₂₀₀ μl of 5 × Kinase buffer (Invitrogen), 5 μl of 1 M MgCl _2, 1 μl of 1 M DTT, 1 μl of 1 M MnCl ₂ and 793 μl of ddH ₂ O

5 × TK substrate-biotin and ATP working fluid: TK substrate - Biotin and ATP concentrations, see Table 1. TK substrate-biotin and ATP were diluted with 1X kinase buffer to 5 times the concentration of the reaction;

5 × Kinase Working Fluid: Concentration of VEGFR2 kinase, see Table 1. The 5 × kinase working fluid was prepared with 1 × Kinase buffer;

4 × Sa-XL665 Working fluid: See Table 1 for the concentration of Sa-XL665 (Cisbio) in the reaction. 4 x Sa-XL665 working fluid was prepared with assay buffer (Cisbio).

4 × TK Ab-cryptite working fluid: TK Ab-cryptate (Cisbio) was diluted 100 times with Assay Buffer (Cisbio) as the working fluid;

2.2 Experimental Procedure

Experimental procedure of HTRF KinEASE TK kit

After all the reagents were prepared as described above, the reagents except for the enzyme were made to have a balance with room temperature, and then samples were added.

 Substrate-biotin, ATP, VEGFR2 kinase and compounds at constant concentrations were reacted in 1X kinase buffer for 20 minutes at room temperature. The concentration for the test compound was 0-100 μM and 2.5% DMSO was used as the co-solvent. To all reaction wells were added 5 μl of 4 × Sa-XL665 working fluid and 5 μl of 4 × TK Ab-cryptite working fluid. After reacting for 1 hour at room temperature, a fluorescence signal (excitation at 320 nm, emission at 665 nm, 615 nm) was detected by an ENVISION detector (PerkinElmer). The inhibition rate for each well was calculated based on the full-active well and the background signal well, and the mean value was used in duplicate wells. Expert software Graphpad PRISM 5.0 was used to fit a half maximal inhibitory concentration for each test compound.

The flow chart of the sample addition is as follows:

2.3 Data Analysis

Emission light ratio (ER) = 665 nm Emission signal / 615 nm Emission signal

Inhibition rate = (ER _positive- ER _sample ) / (ER _positive- ER _negative ) * 100%

3. Experimental Results

The IC ₅₀ values of the compounds for kinase VEGFR2 were determined using the HTRF KinEASE TK kit. The final concentrations of the compounds starting from 100 [mu] M and 4 fold dilution of the gradient were measured to provide 10 concentrations. Replicate wells were used for each concentration. The final concentration of DMSO was adjusted to 1% in the reaction system. The experimental results are shown in Table 2 and Fig.

Table 2 IC ₅₀ determinations of compounds of the invention for inhibitory activity of VEGFR2 kinase

4. Experimental Results:

All of the compounds according to the invention according to the invention have an IC ₅₀ value of less than 1000, indicating that the compounds according to the invention are excellent in inhibiting activity of kinase VEGFR2 and can be studied as excellent anticancer agents.

Compared with the superior anticancer drug Sorapenib on the market, the preferred compound of the present invention is 2-3 times higher than the commercially available drug Sorapenib (comparative compound 1) in view of the inhibitory activity of the kinase VEGFR2, and is disclosed in Chinese patent application CN201110435847.9 Was better than the disclosed compound (Comparative Compound 2). The IC ₅₀ values of the compounds prepared in Example 1 of the present invention were 0.32 and 0.15 times the Soraphenib (free base) and Comparative Compound 2, respectively, i.e., their inhibitory activities were 3.13 times and 6. 3 It was a ship. The IC ₅₀ values of the compounds prepared in Example 2 were 0.62 and 0.34, respectively, of soraphenyl (free base) and Comparative Compound 2, that is, their inhibitory activities were 1.61 and 3.4 times of sorapenib and Comparative Compound 2, respectively . The IC ₅₀ values of the compounds prepared in Example 4 were 0.75 and 0.36 times of soraphenyl (free base) and Comparative Compound 2, respectively, that is, their inhibitory activities were 1.3 times and 2.7 times of Sorapanib and Comparative Compound 2, respectively. The IC ₅₀ value of the compound prepared in Example 3 was comparable to that of Comparative Compound 2 at 2.5 times that of sorapenib (free base), i.e. its inhibitory activity was 1.5 times that of sorapenib, i.e. its inhibitory activity was comparable Respectively.

Thus, from the above experimental results, it can be seen that the compound according to the present invention has excellent inhibitory activity of VEGFR2 kinase.

By comparison of X3, X4 and X5 in the general formula II of the present invention, the compounds according to the invention are clearly better than those of Comparative Examples 3, 4 and 5.

Experimental Example 2: Determination of the compound according to the present invention against the IC ₅₀ of in vitro anti-proliferation of tumor cells

1. Materials and Methods

Cell strain:

MDA-MB-231 human breast cancer cell line (purchased from Shanghai Institute of Cell Biology, Chinese Academy of Sciences);

A498 human kidney cancer cell line (purchased from Shanghai Institute of Cell Biology, Chinese Academy of Sciences);

HCT116 human colon cancer cell line (purchased from Shanghai Institute of Cell Biology, Chinese Academy of Sciences);

786-O human clear cell carcinoma cell line (purchased from Shanghai Institute of Cell Biology, Chinese Academy of Sciences);

MiaPaCa-2 human pancreatic cancer cell line (purchased from American ATCC);

SK-OV-3 human ovarian cancer cell line (purchased from Shanghai Institute of Cell Biology, Chinese Academy of Sciences);

HepG2 human liver cancer cell line (purchased from Shanghai Institute of Cell Biology, Chinese Academy of Sciences);

NCI-H460 human large cell lung cancer cell line (purchased from Shanghai Institute of Cell Biology, Chinese Academy of Sciences);

HL-60 human acute myelogenous leukemia cell line (purchased from Shanghai Institute of Cell Biology, Chinese Academy of Sciences);

Reagents and consumables:

Cell Counting Kit-8 (Cat # CK04-13, Dojindo);

96-well plates (Cat # 3599, Corning Costar);

Fetal bovine serum (Cat # 10099-141, GIBCO);

Culture media (Invitrogen) in Table 3;

Desk top ELISA instrument Spectra Max M5 Microplate Reader (Molecular Devices);

Test compound: The compound prepared in Example

Control Compound: Compound prepared in Comparative Example

2. Experimental steps

2.1 Reagent preparation

Table 3 Preparation of culture medium

Preparation of the compound: The compound was diluted to a final concentration of 10 mM using DMSO.

2.2 Cell culture

a) The cells of the logarithmic growth phase were recovered, counted and re-suspended in complete medium.

b) The cell concentration was adjusted to the appropriate concentration and the cells were inoculated in 96-well plates in 100 μl cell suspension per well.

c) Cells were cultured in an incubator at 37 ° C, 100% relative humidity, 5% CO ₂ for 24 hours.

2.3 IC ₅₀ Assay

a) Cells from logarithmic growth phase were recovered, counted and resuspended in complete medium. The concentration of the cells was adjusted to an appropriate concentration (determined according to the optimal experimental results of cell density) and the cells were inoculated into 96-well plates with 100 μl of cell suspension per well. The cells were incubated in an incubator at 37 ° C, 100% relative humidity, 5% CO ₂ for 24 hours.

b) diluting the test compound with 500 [mu] M using the culture medium, followed by 8 dilutions of the gradient. Cells were added at 25 [mu] l / well. The final concentration of the compound was diluted 4-fold in the range of 100 [mu] M and 0 [mu] M, including 10 concentrations.

c) Cells were cultured in an incubator at 37 ° C, 100% relative humidity, 5% CO ₂ for 72 hours.

d) The culture medium was taken out, discarded, complete medium containing 10% CCK-8 was added, and the cells were then incubated in the incubator at 37 ° C for 2-4 hours.

e) After slight shaking, the absorbance at 450 nm wavelength was determined using a SprctraMax M5 Microplate Reader and the inhibition rate was calculated using the absorbance at 650 nm wavelength as a reference.

2.4 Data Processing

The tumor cell proliferation inhibition rate of the drug is calculated by the following equation:

Tumor cell proliferation inhibition rate (%) = [(A _c -A _s ) / (A _c -A _b ) x 100%

A: absorbance of the sample (cell + CCK-8 + test compound);

A _c : absorbance of negative control (Cell + CCK-8 + DMSO);

A _b : absorbance of positive control (culture medium + CCK-8 + DMSO);

IC ₅₀ curves were fitted and IC ₅₀ values were calculated using the specialized software GraphPad Prism 5.0.

3. Experimental Results

In the experiment, compounds according to the invention were determined for IC ₅₀ values of in vitro anti-proliferation of tumor cell lines. The final concentration of the compound was diluted 4-fold in the range of 100 [mu] M and 0 [mu] Md containing 10 concentrations.

The experimental results are shown in Table 4.

Table 4 IC ₅₀ (μM)

4. Experimental Conclusion:

In an experiment on the antiproliferation of tumor cells in vitro, the compound according to the present invention had an IC ₅₀ of 50% inhibitory concentration of 0 to 20, indicating that the compounds according to the present invention have excellent inhibitory activity against tumor cells in vitro and excellent anti- As a result.

Compared to sorafenib, a very superior anticancer drug on the market, the compounds according to the present invention have half-maximal inhibitory concentrations of different tumor cell lines such as SK-OV-3, HCT-116, 786-O and MDA-MB- inhibitor concentration, IC ₅₀ ) (for example, for MDA-MB-231, the IC ₅₀ of the compound prepared in Example 1 was 0.28-fold higher than that of sorapenib , And the IC ₅₀ of the compound prepared in Example 2 was 0.3 times that of sorapenib); The half-maximal inhibitory concentrations of tumor cell lines such as A498, MiaPaCa-2, HepG2, NCI-H460 and HL-60 were comparable to the commercially available drug sorapenib.

This experiment demonstrates that the compounds according to the present invention are excellent in anti-proliferative activity of tumor cells.

Experimental Example 3: Study on the pharmacokinetics of the compounds according to the present invention in mice

1. Materials and Methods

1.1 Test compound

In Examples of the present invention and Comparative Example,

1.2 Experimental animals

CD-1 mice, female, weighing 28-35 g.

1.3 Routes of administration

Routes of administration: Intravenous injection (IV); Per os (PO)

Fasting conditions: You can drink water freely, and there is no fasting.

2. Experimental Method

2.1 Administration and sample collection

2.1.1 Administration

The mice were weighed prior to administration and the dose volume was calculated based on their body weight (IV group: 4 mL / kg; PO group: 10 mL / kg).

Routes of administration and dose: intravenous (IV) group: 1 mg / kg; Oral (PO) group: 5 mg / kg).

Sample: Plasma.

Animal grouping: 3 mice / group, IV group and PO group for each test compound

2.1.2 Sampling

After administration, mice were sacrificed at the pre-determined time points (5 min, 15 min, 30 min, 1 hr, 2 hr, 4 hr, 8 hr, 24 hr) 30 袖 L whole blood was collected and 30 袖 L whole blood was collected from the rim of the mice in the PO group at predetermined time points (15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, 24 hours) Respectively. Whole blood was centrifuged (6,000 rpm, 5 minutes) to obtain plasma. All plasma samples were stored in a -80 ° C refrigerator for further analysis.

2.2 Quantitative analysis method

Conditions for LC / MS / MS are as follows:

Ionization mode: ESI, cation;

Detection mode: MRM;

Quantitative ion FD2012015: 506.12 / 270.20; Internal standard (terpenadine): 472.40 / 436.40;

Sample treatment: Protein was precipitated using 50 ng of terpenazine solution in acetonitrile solution;

Samples: CD-1 mouse plasma (using EDTA to prevent blood clotting);

Sample volume: 20 mu L;

Chromatographic column: ACE C4 column (50 mm * 2.1 mm, 5 micron)

Moble phase: gradient elution, mobile phase A was water (containing 0.1% formic acid) and mobile phase B was acetonitrile (containing 0.1% formic acid);

Flow rate (mL / min): 0.9;

Column temperature (占 폚): room temperature;

Injection volume (μL): 5;

Time in minutes: 2.0.

3. Data processing

Pharmacokinetic parameters were evaluated according to a non-compartment model (calculated by WinNonlin software).

IV _{Parameters: t 1/2 (hr)} ; C ₀ (ng / mL); AUC _last (hr * ng / mL); AUC _Inf (hr * ng / mL); AUC Extr (%); Vz (L / kg); Vss (L / kg); CL (mL / min / kg); MRT (hr).

Parameter _{PO: t 1/2 (hr)} ; t _max (hr); C _max (ng / mL); AUC _last (hr * ng / mL); AUC _Inf (hr * ng / mL); AUC ExTr (%); MRT (hr); AUC / D (hr * mg / mL); F (%).

4. Experimental Results:

Table 5 Pharmacokinetic parameters of the compounds according to the invention after oral administration

     (Mean value from 3 mice per group)

Note: Cmax: peak concentration; AUC _last : area under curve; F: Oral bioavailability

5. Experimental Conclusion:

The compounds of Examples 1 and 2 were significantly superior to sorafenib, which is commercially available in terms of metabolic stability, peak concentration, area under the curve of concentration-time, and oral bioavailability, and therefore the prospects for clinical application are very good. This is because, after introduction of a substituent at the X3 position of the general formula I, the substituent (which is hydrogen at the X3 position of the comparative compound 2, 3 or 4) easily blocks the site of metabolism, To improve the metabolic stability and to ensure the high blood concentration of the compound in vivo, thereby further enhancing the efficacy of the compounds according to the present invention.

Experimental Example 4

1. Cell culture

786-O cells were cultured in RPMI-1640 culture medium containing 2 mM glutamine in addition to inactivated 10% fetal bovine serum, 100 U / ml penicillin and 100 ug / mL streptomycin in an incubator at 37 캜 and 5% CO ₂ . During cell culture, the initial concentration was 5 x 10 ⁵ cells / ml and the cells were separated into other bottles every 3-4 days until the cells reached 100% cell density. Tumor cells in the logarithmic growth phase were used for in vivo tumor inoculation.

2. Inoculation and classification of tumor cells

Female SCID-Beige hairless nude mice were resuspended in the right lateral thorax using 786-O tumor cells resuspended at 8 × 10 ⁶ cells / 0.1 ml in PBS. . When the tumor grew to a volume of 800 mm ³ , the tumor was removed under aseptic conditions. Fully grown tumor tissue was cut into 2 × 2 × 2 mm ³ tumor masses, which were then inoculated into the animal by subcutaneous injection. When tumors grew to a volume of about 100 mm ³ , mice were divided into groups and administered to all 5 groups, 8 mice per group.

3. Tumor measurement and experimental indexes (experimental indexes)

The volume of the tumor was measured twice weekly using a Vernier caliper. The long and short diameters of the tumor were measured. The volume was calculated by the general formula: volume = 0.5 x long diameter x short diameter ² . After the final measurement, the animals are slaughtered and the tumor removed to weigh. Based on the tumor weight of each group, the tumor growth-inhibiting rate (TGI) was calculated. Tumor growth-inhibition ratio (TGI) = (1-T / C) x 100%, where T represents the mean tumor weight of the drug-administered group; C represents the mean tumor weight of the negative control. Data were statistically analyzed by TGI ≥ 60% and were effective at p <0.05.

Table 6 Tumor-inhibitory effect of FD-2013018 on xenograft tumor-bearing mice (tumor weight) xenografted with humanized renal clear cells 786-O

Notes: ^a. Mean value ± standard error; ^b. Comparison with solvent control; ^c Blank solvent: Solvent DMA: Solutol HS-15: H ₂ O = 5: 5: 90 (by volume) (DMA is dimethylacetamide, Solutol HS-15 is purchased from BASF, CAS: 61909-81-7); FD-2013018 is in free form; po means per os, and QD means once a day.

P: Group 2 (10 mg / kg) to Group 3 (20 mg / kg) = 0.999; Group 2 vs. Group 4 (40 mg / kg) = 0.386; Group 3 vs. Group 4 = 0.270.

Experimental Example 5

1. Cell culture

HCT116 cells were cultured in a McCoy'5 5a culture medium containing 2 mM glutamine in addition to inactivated 10% fetal bovine serum, 100 U / ml penicillin and 100 ug / mL streptomycin in an incubator at 37 캜 and 5% CO ₂ . During cell culture, the initial concentration was 5 x 10 ⁵ cells / ml and the cells were separated into other bottles every 3-4 days until the cells reached 100% cell density. Tumor cells in the logarithmic growth phase were used for in vivo tumor inoculation.

2. Inoculation and classification of tumor cells

Using the re-suspended the HCT116 tumor cells in 1.0 × 10 ⁷ cells / 0.1ml in PBS condition in Balb / c hairless mice, were inoculated by subcutaneous injection in the right lateral chest. When the tumor grew to a volume of 800 mm ³ , the animal was sacrificed and the tumor was removed under aseptic conditions. Fully grown tumor tissue was cut into 2 × 2 × 2 mm ³ sized tumor mass, which was then used to inoculate the animal by subcutaneous injection in the right lateral chest. When tumors grew to a volume of about 110 mm ³ , mice were divided into groups and administered to all 5 groups, 8 mice per group.

3. Measurement and experimental index of tumor

Using a vernier caliper, the volume of the tumor was measured twice a week. The long and short diameters of the tumors were measured. The volume was calculated by the general formula: volume = 0.5 x long diameter x short diameter ² . After the final measurement, the animals are slaughtered and the tumor removed to weigh. Based on the tumor weight, the tumor growth-inhibition ratio (TGI) was calculated. Tumor growth-inhibition ratio (TGI) = (1-T / C) x 100%, where T represents the average tumor weight of the test compound group; C represents the average tumor weight of the solvent control. When the assay was terminated, the experimental animals were euthanized.

Table 7 [0072] On mice (tumor weight) with tumor xenografts transfected with HCT1166

     Tumor-inhibiting effect of FD-2013018

Notes: ^a. Mean value ± standard error; ^b. Comparison with control group; FD-2013018 was in free form; The blank solvent is the same as defined in Table 6;

P: Group 2 (10 mg / kg) to Group 3 (20 mg / kg) = 0.687; Group 2 vs. Group 4 (40 mg / kg) = 0.248;

Group 3 vs. Group 4 = 0.807.

Comparative Experimental Example 1:

Sunitinib was prepared by the method disclosed in patent WO0160814A1.

Preparation of animal models:

A fully grown 786-O solid tumor was cut under sterile conditions into a mass of about 1 mm ³ of an average volume, which was used to inoculate the hairless mouse in the axillary cavity of the right forearm armpit. Tumor growth was routinely observed until the tumors grew to 250-550 mm ³ .

Classification and administration:

Animals with too large or small, irregularly shaped tumors were discarded. A total of 48 mice with tumors with tumor volumes of 250-550 mm ³ in good condition were divided into 6 groups: 1 solvent control, 3 positive controls, and 2 test samples. Administered intragastrically once a day for both positive control and test sample groups; The solvent control group was administered once per day with 12.5% ethanol and 12.5% water solution of polyoxyethylene castor oil (caster oil); The intragastrical volume was 10 mL / kg.

During the dosing period, tumor diameters were measured twice a week, tumor volume was calculated, and animal weights were recorded. Animal status was observed and the abnormal state recorded until administration.

Animal slaughter:

The animals were slaughtered with CO ₂ , the tumors were removed, weighed and photographed. The animals were visually anatomically treated, and organs were observed by visual observation.

Observations:

Inhibition of tumor weight (IR) = (W _C -W _T ) / W _C

Here, W _C And W _T were the mean tumor weight of the solvent control group and the average tumor weight of the drug administration group, respectively.

BW ₀ represents the body weight (ie, d ₀ ) of an animal weighed at the time of classification, and BW _t represents the weight of an animal weighed at all times. If the weight loss rate is negative, it means that the weight is increased.

Statistical methods:

We used Microsoft Office Excel 2003 software to perform computation and statistical processing on experimental data. Unless otherwise specified, data are expressed as mean ± standard error (Mwan ± Standard Error, mean ± SE) and t-test is used for comparison between two groups.

&Lt; tb &gt; &lt; tb &gt; &lt; tb &gt; &lt; SEP &gt; 8 &lt; SEP &gt;

     Effect of FD-1210005 on tumor weight

Note: 1. Compared to solvent control, ^* P <0.05, ^** P <0.01;

2. In the experiment, sorapenib was present in the form of p-toluene sulfonate and FD-1210005 in free form.

Comparative Experimental Example 2

Preparation of animal models:

Using a human HCT-116 cell suspension sufficiently grown in logarithmic growth under aseptic conditions, hairless mice were inoculated subcutaneously through the hollow portion of the right foreleg axilla via an injector. Tumor growth was routinely monitored until tumors were grown to a volume of 100-300 mm ³ .

Classification and administration:

Forty-eight mice with tumors ranging in volume from 100 to ³⁰⁰ mm ³ were selected and divided into six groups, one solvent control group, three positive control groups and two test sample groups. Once daily for both positive control and test sample groups; The solvent control group was administered once per day with 12.5% ethanol and 12.5% water solution of polyoxyethylene castor oil; The stomach volume was 10 mL / kg.

During the dosing period, tumor diameters were measured twice a week, tumor volume was calculated, and animal weights were recorded. Animal status was observed until admission and abnormalities were recorded.

Animal slaughter:

The animals were slaughtered with CO ₂ , the tumors were removed, weighed and photographed. After the photographs were taken, each tumor mass was cut into two parts, one was stored in 4% paraformaldehyde and the other was packed in a freezing tube and frozen with liquefied nitrogen. Animals were visually anatomically treated and organs were observed with eyes to confirm their normal state.

Observations:

Inhibition of tumor weight (IR) = (W _C -W _T ) / W _C

Here, W _C And W _T were the mean tumor weight of the solvent control group and the average tumor weight of the drug administration group, respectively.

Statistical methods:

We used Microsoft Office Excel 2003 software to perform computation and statistical processing on experimental data. Unless otherwise specified, the data are expressed as mean ± standard error (mean ± SE) and the t-test is used for comparison between the two groups.

&Lt; tb &gt; &lt; tb &gt; &lt; SEP &gt;

     Effect of FD-1210005 on

Note: 1. Compared to solvent control, ^* P <0.05, ^** P <0.01;

    2. "-" indicates no data.

    3. In the experiment, sorapenib was present in the form of p-toluenesulfonate

And FD-1210005 existed in free form.

In vivo antitumor ( in The results of the vivo antitumor experiments show that the compound (FD-2013018) prepared in Example 2 was 82% and 84.1% at a dose of 10 mg / kg for mice with tumor xenotransplanted with 786-O and HCT116, Of tumor-inhibiting effect, reaching a tumor-inhibiting effect of 92% at a dose of 40 mg / kg; The tumor-inhibiting effect of 80% and 67% was reached at a dose of 20 mg / kg for mice with tumor xenografts in which Comparative Compound 2 (FD-2010005) was xenotransplanted with 786-O and HCT116, respectively, Lt; RTI ID = 0.0 &gt; 80% &lt; / RTI &gt; and 71% tumor-inhibiting effect. The data show that the compound (FD-2013018) prepared in Example 2 was superior to Comparative Compound 2 (FD-2010005) in terms of antitumor activity in vivo and had a lower effective dose and a stronger anti- &Lt; / RTI &gt; activity.

In summary, the compounds according to the present invention have a very strong antitumor activity in vitro and in vivo, and are particularly excellent in pharmacokinetic properties. Compared with sorapenib and the compound of Comparative Example 2, the compounds according to the present invention have more potent anti-tumor activity and better pharmacokinetic properties in vitro and in vivo.

Having described the embodiments of the present invention in detail, those skilled in the art will recognize that changes and substitutions can be made in accordance with the teachings of the present invention, and all such changes and modifications are within the scope of protection of the present invention will be. The scope of the invention is defined by the claims and any equivalents thereof.

Claims (40)

A multisubstituted pyridine compound of formula I, or a hydrate, solvate or pharmaceutically acceptable salt thereof:

Wherein X &lt; ₁ &gt; is selected from substituted or unsubstituted 5 membered heteroaromatic rings of formula a;

R ₄ , R ₅ and R ₆ are each independently selected from the group consisting of a carbon atom, a nitrogen atom, an oxygen atom or a sulfur atom, and R ₈ , R ₉ and R ₁₀ are each independently selected from the group consisting of hydrogen, halogen, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxyl;
X ₂ is selected from the group consisting of F and H;
X ₃ is halogen, -CN, C ₁ -C ₄ alkyl, halogenated C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxyl, halogenated C ₁ -C ₄ An alkoxyl, and -NR ₁₁ R _12, R ₁₁ And R ₁₂ are each independently selected from the group consisting of hydrogen and C ₁ -C ₄ alkyl. The method according to claim 1,
R ₄ , R ₅ and R ₆ are each independently selected from the group consisting of carbon atoms and nitrogen atoms, or a hydrate, solvate or pharmaceutically acceptable salt thereof. The method according to claim 1 or 2,
R ₄ , R ₅ and R ₆ are not simultaneously carbon atoms, or a hydrate, solvate or pharmaceutically acceptable salt thereof. The method according to claim 1 or 2,
R ₄ , R ₅ and R ₆ are not simultaneously nitrogen atoms, or a hydrate, solvate or pharmaceutically acceptable salt thereof. 5. The method according to any one of claims 1 to 4,
R ₈ , R ₉ and R ₁₀ are each independently selected from the group consisting of hydrogen and methyl, or a hydrate, solvate or pharmaceutically acceptable salt thereof. The method according to claim 1,
X ₁ is

, A multisubstituted pyridine compound, or a hydrate, solvate, or pharmaceutically acceptable salt thereof. 7. The method according to any one of claims 1 to 6,
X ₃ is selected from the group consisting of F, Cl, Br, -CF ₃ , -CN, C ₁ -C ₂ alkyl, C ₁ -C ₂ alkoxyl and -NR ₁₁ R ₁₂ ; R ₁₁ and R ₁₂ are each hydrogen and C ₁ -C ₂ is independently selected from the group consisting of alkyl, a multi-substituted pyridine compound, or a hydrate thereof, acceptable salt solvate or pharmaceutically. The method of claim 7,
X ₃ is selected from the group consisting of F, Cl and -CN, or a hydrate, solvate or pharmaceutically acceptable salt thereof. The method according to claim 1,
A multisubstituted pyridine compound of general formula I is selected from the group consisting of the following compounds: or a hydrate, solvate or pharmaceutically acceptable salt thereof.

,

,

,

,

,

,

And

. The method according to claim 1,
A multisubstituted pyridine compound of general formula I is selected from the group consisting of the following compounds: or a hydrate, solvate or pharmaceutically acceptable salt thereof.

, And

. The method according to any one of claims 1 to 10,
Pharmaceutically acceptable salts of the polyunsaturated pyridine compounds of the general formula I are the hydrochloride, hydrobromide, sulfate, phosphate, methanesulfonate, trifluoromethanesulfonate, benzenesulfonate, p-toluenesulfonate, 1- Naphthalene sulfonate, 2-naphthalene sulfonate, acetate, trifluoroacetate, malate, tartarate, citrate, lactate, oxalate, succinate, fumarate, maleate, benzoate, salicylate, phenyl A substituted or unsubstituted pyridine compound selected from the group consisting of an acetic acid salt and a mandelic acid salt, or a hydrate, solvate or pharmaceutically acceptable salt thereof. A pharmaceutical composition comprising a polysubstituted pyridine compound according to any one of claims 1 to 10, or a hydrate, solvate or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable adjuvant. A process for the preparation of a polysubstituted pyridine compound according to any one of claims 1 to 10, comprising the steps of:
1) reacting a compound of formula B and a compound of formula C in the presence of potassium tert-butoxide as a catalyst to give a compound of formula D, as shown by the following scheme:

,
Wherein R &lt; ₁₃ &gt; is F, Cl, Br or I;
2) The compound of formula D and the compound of formula E are reacted in the presence of tetrakis (triphenylphosphine) palladium or bis (triphenylphosphine) palladium (II) dichloride as catalyst, Lt; / RTI &gt; to obtain a compound of general formula F:

; And
3) reacting a compound of formula F with a compound of formula G to yield a polysubstituted pyridine compound of formula I, as shown by the following scheme:

;
Wherein X ₁ , X ₂ , and X ₃ have the same meanings as defined in any one of claims 1 to 10. 14. The method of claim 13,
The compound of formula B is prepared by the following method:
Halogenating the compound of formula A to give the compound of formula B:

Wherein R ₁₃ is F, Cl, Br or I. The method according to any one of claims 1 to 10,
Wherein X &lt; ₃ &gt; is NH &lt; _{2 &} gt ;, said method comprising the steps of:
1) reacting a compound of general formula H and a compound of general formula C as bases in the presence of potassium tert-butoxide to give a compound of general formula W, as shown by the following scheme:

;
Wherein R &lt; ₁₃ &gt; is F, Cl, Br or I;
2) The compound of formula W and the compound of formula E are reacted in the presence of tetrakis (triphenylphosphine) palladium or bis (triphenylphosphine) palladium (II) dichloride as catalyst, To obtain a compound of the general formula J:

;
3) hydrogenating the compound of formula J in the presence of palladium-carbon as catalyst to give the compound of formula K

; And
4) reacting a compound of formula K with a compound of formula G to give a compound of formula L, as shown in the following scheme:

A pharmaceutical composition for the treatment and / or prophylaxis of diseases associated with VEGFR-2, VEGFR-3, CRAF, PDGFR-β, BRAF, V600E BRAF, KIT and / Or a hydrate, solvate or pharmaceutically acceptable salt thereof. 18. The method of claim 16,
Wherein said disease associated with VEGFR-2, VEGFR-3, CRAF, PDGFR-beta, BRAF, V600E BRAF, KIT and / or FLT-3 kinase comprises a tumor or cancer. 18. The method of claim 17,
Wherein said tumor or cancer is selected from the group consisting of malignant melanoma, liver cancer, renal cancer, acute leukemia, chronic leukemia, non-small cell lung cancer, prostate cancer, thyroid cancer, skin cancer, colon cancer, rectal cancer, pancreatic cancer, ovarian cancer, Or mesothelioma. A method for the treatment and / or prophylaxis of a disease associated with VEGFR-2, VEGFR-3, CRAF, PDGFR-beta, BRAF, V600E BRAF, KIT and / or FLT-3 kinase, Comprising administering to a subject a therapeutically or prophylactically effective amount of a multisubstituted pyridine compound according to one of the preceding claims, or a hydrate, solvate, or pharmaceutically acceptable salt thereof. The method of claim 19,
Wherein said disease associated with VEGFR-2, VEGFR-3, CRAF, PDGFR-beta, BRAF, V600E BRAF, KIT and / or FLT-3 kinase comprises a tumor or cancer. The method of claim 20,
Wherein said tumor or cancer is selected from the group consisting of malignant melanoma, liver cancer, renal cancer, acute leukemia, chronic leukemia, non-small cell lung cancer, prostate cancer, thyroid cancer, skin cancer, colon cancer, rectal cancer, pancreatic cancer, ovarian cancer, Or mesothelioma. The use of any one of claims 1 to 10 for use in the treatment and / or prevention of diseases associated with VEGFR-2, VEGFR-3, CRAF, PDGFR-β, BRAF, V600E BRAF, KIT and / Or a hydrate, solvate and pharmaceutically acceptable salt thereof. 23. The method of claim 22,
Such diseases associated with VEGFR-2, VEGFR-3, CRAF, PDGFR-beta, BRAF, V600E BRAF, KIT and / or FLT-3 kinase are multisubstituted pyridine compounds, or hydrates, solvates and pharmacology thereof, Acceptable salt. 24. The method of claim 23,
Wherein said tumor or cancer is selected from the group consisting of malignant melanoma, liver cancer, renal cancer, acute leukemia, chronic leukemia, non-small cell lung cancer, prostate cancer, thyroid cancer, skin cancer, colon cancer, rectal cancer, pancreatic cancer, ovarian cancer, Or mesothelioma, or a hydrate, solvate and pharmaceutically acceptable salt thereof. A method for inhibiting the activity of VEGFR-2, VEGFR-3, CRAF, PDGFR-beta, BRAF, V600E BRAF, KIT and / or FLT-3 kinase in a cell, Comprising administering to said cell an effective amount of a pyridine compound, or a hydrate, solvate, and a pharmaceutically acceptable salt thereof. 26. The method of claim 25,
The method may be used in vitro . 26. The method of claim 25,
Wherein the method is carried out in vivo. 26. A method according to any one of claims 25-27,
Wherein the cell is a cell line, or a cell derived from an object such as a tumor cell or a cancer cell. 29. The method of claim 28,
Wherein said tumor or cancer is selected from the group consisting of malignant melanoma, liver cancer, renal cancer, acute leukemia, chronic leukemia, non-small cell lung cancer, prostate cancer, thyroid cancer, skin cancer, colon cancer, rectal cancer, pancreatic cancer, ovarian cancer, &Lt; / RTI &gt; and mesothelioma. The use of a multisubstituted pyridine compound according to any one of claims 1 to 10, or a hydrate, solvate or pharmaceutically acceptable salt thereof in the manufacture of a medicament, wherein said medicament is capable of inhibiting VEGFR-2, VEGFR-3 , CRAF, PDGFR-β, BRAF, V600E BRAF, KIT and / or FLT-3 kinase. 32. The method of claim 30,
Wherein said medicament is used in an in vitro method. 32. The method of claim 30,
Wherein said medicament is used in an in vivo method. 32. The method according to any one of claims 30 to 32,
Wherein said cell is a cell derived from a subject such as a cell line or a tumor cell or cancer cell. 34. The method of claim 33,
Wherein said tumor or cancer is selected from the group consisting of malignant melanoma, liver cancer, renal cancer, acute leukemia, chronic leukemia, non-small cell lung cancer, prostate cancer, thyroid cancer, skin cancer, colon cancer, rectal cancer, pancreatic cancer, ovarian cancer, And mesothelioma. Use of a compound according to any one of claims 1 to 10 for use in inhibiting the activity of VEGFR-2, VEGFR-3, CRAF, PDGFR-beta, BRAF, V600E BRAF, KIT and / or FLT- Substituted pyridine compounds, or hydrates, solvates, and pharmaceutically acceptable salts thereof. 36. The method of claim 35,
A polysubstituted pyridine compound, or a hydrate, solvate, and pharmaceutically acceptable salt thereof, for use in an in vitro method. 36. The method of claim 35,
A multisubstituted pyridine compound, or a hydrate, solvate, and pharmaceutically acceptable salt thereof, for use in an in vivo method. 35. The method according to any one of claims 35 to 37,
Wherein said cell is a cell derived from an object such as a cell line or a tumor cell or cancer cell, or a hydrate, solvate, or pharmaceutically acceptable salt thereof. 42. The method of claim 38,
Wherein said tumor or cancer is selected from the group consisting of malignant melanoma, liver cancer, renal cancer, acute leukemia, chronic leukemia, non-small cell lung cancer, prostate cancer, thyroid cancer, skin cancer, colon cancer, rectal cancer, pancreatic cancer, ovarian cancer, And mesothelioma, or a hydrate, solvate, and pharmaceutically acceptable salt thereof. A kit for inhibiting the activity of VEGFR-2, VEGFR-3, CRAF, PDGFR-β, BRAF, V600E BRAF, KIT and / or FLT-3 kinase in a cell, Or a hydrate, solvate or pharmaceutically acceptable salt thereof, and, optionally, instructions.

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